Gene ontology annotations for ADAM9
Experiment description of studies that identified ADAM9 in exosomes
1
Experiment ID
489
MISEV standards
✔
EM
EV Biophysical techniques
✔
Cd9|Cd81|Cd63|Gapdh|Sdcbp|Lamp1|Aqp1|Rab5a|Icam1|Cd82|Itga2b|Tsg101|Lamp2|Rab35|Flot1|Flot2|Cd151|Rab5b|Tfrc|Uchl1
EV Enriched markers
✔
Canx
EV Negative markers
✔
NTA
EV Particle analysis
Identified molecule
Protein
Identification method
Mass spectrometry
PubMed ID
36408942
Organism
Rattus norvegicus
Experiment description
Quantitative proteomic analysis of exosomes from umbilical cord mesenchymal stem cells and rat bone marrow stem cells
Authors
Xu X, Yin F, Guo M, Gan G, Lin G, Wen C, Wang J, Song P, Wang J, Qi ZQ, Zhong CQ.
Journal name
Proteomics
Publication year
2023
Sample
Bone marrow mesenchymal stem cells
Sample name
BMSC - Passage 6
Isolation/purification methods
Differential centrifugation Filtration Ultracentrifugation
Flotation density
-
Molecules identified in the study
Protein
Methods used in the study
Western blotting Mass spectometry
2
Experiment ID
490
MISEV standards
✔
EM
EV Biophysical techniques
✔
Cd9|Cd81|Cd63|Gapdh|Sdcbp|Lamp1|Aqp1|Rab5a|Icam1|Cd82|Itga2b|Tsg101|Lamp2|Rab35|Flot1|Flot2|Cd151|Rab5b|Tfrc|Uchl1
EV Enriched markers
✔
Canx
EV Negative markers
✔
NTA
EV Particle analysis
Identified molecule
Protein
Identification method
Mass spectrometry
PubMed ID
36408942
Organism
Rattus norvegicus
Experiment description
Quantitative proteomic analysis of exosomes from umbilical cord mesenchymal stem cells and rat bone marrow stem cells
Authors
Xu X, Yin F, Guo M, Gan G, Lin G, Wen C, Wang J, Song P, Wang J, Qi ZQ, Zhong CQ.
Journal name
Proteomics
Publication year
2023
Sample
Bone marrow mesenchymal stem cells
Sample name
BMSC - Passage 7
Isolation/purification methods
Differential centrifugation Filtration Ultracentrifugation
Flotation density
-
Molecules identified in the study
Protein
Methods used in the study
Western blotting Mass spectometry
3
Experiment ID
491
MISEV standards
✔
EM
EV Biophysical techniques
✔
Cd9|Cd81|Cd63|Gapdh|Sdcbp|Lamp1|Aqp1|Rab5a|Icam1|Cd82|Itga2b|Tsg101|Lamp2|Rab35|Flot1|Flot2|Cd151|Rab5b|Tfrc|Uchl1
EV Enriched markers
✔
Canx
EV Negative markers
✔
NTA
EV Particle analysis
Identified molecule
Protein
Identification method
Mass spectrometry
PubMed ID
36408942
Organism
Rattus norvegicus
Experiment description
Quantitative proteomic analysis of exosomes from umbilical cord mesenchymal stem cells and rat bone marrow stem cells
Authors
Xu X, Yin F, Guo M, Gan G, Lin G, Wen C, Wang J, Song P, Wang J, Qi ZQ, Zhong CQ.
Journal name
Proteomics
Publication year
2023
Sample
Bone marrow mesenchymal stem cells
Sample name
BMSC - Passage 8
Isolation/purification methods
Differential centrifugation Filtration Ultracentrifugation
Flotation density
-
Molecules identified in the study
Protein
Methods used in the study
Western blotting Mass spectometry
4
Experiment ID
492
MISEV standards
✔
EM
EV Biophysical techniques
✔
Cd9|Cd81|Cd63|Gapdh|Sdcbp|Lamp1|Aqp1|Rab5a|Icam1|Cd82|Itga2b|Tsg101|Lamp2|Rab35|Flot1|Flot2|Cd151|Rab5b|Tfrc|Uchl1
EV Enriched markers
✔
Canx
EV Negative markers
✔
NTA
EV Particle analysis
Identified molecule
Protein
Identification method
Mass spectrometry
PubMed ID
36408942
Organism
Rattus norvegicus
Experiment description
Quantitative proteomic analysis of exosomes from umbilical cord mesenchymal stem cells and rat bone marrow stem cells
Authors
Xu X, Yin F, Guo M, Gan G, Lin G, Wen C, Wang J, Song P, Wang J, Qi ZQ, Zhong CQ.
Journal name
Proteomics
Publication year
2023
Sample
Bone marrow mesenchymal stem cells
Sample name
BMSC - Passage 9
Isolation/purification methods
Differential centrifugation Filtration Ultracentrifugation
Flotation density
-
Molecules identified in the study
Protein
Methods used in the study
Western blotting Mass spectometry
5
Experiment ID
207
MISEV standards
✔
EM
EV Biophysical techniques
✔
TSG101|HSP70|FLOT1
EV Enriched markers
✔
VDAC
EV Negative markers
✔
NTA
EV Particle analysis
Identified molecule
protein
Identification method
Mass spectrometry
PubMed ID
23161513
Organism
Homo sapiens
Experiment description
Proteomic analysis of exosomes from mutant KRAS colon cancer cells identifies intercellular transfer of mutant KRAS.
Authors
Demory Beckler M, Higginbotham JN, Franklin JL, Ham AJ, Halvey PJ, Imasuen IE, Whitwell C, Li M, Liebler DC, Coffey RJ.
Journal name
Mol Cell Proteomics
Publication year
2012
Sample
Colorectal cancer cells
Sample name
DKO-1
Isolation/purification methods
Differential centrifugation Filtration
Flotation density
-
Molecules identified in the study
Protein
Methods used in the study
Mass spectrometry
6
Experiment ID
208
MISEV standards
✔
EM
EV Biophysical techniques
✔
TSG101|HSP70|FLOT1
EV Enriched markers
✔
VDAC
EV Negative markers
✔
NTA
EV Particle analysis
Identified molecule
protein
Identification method
Mass spectrometry
PubMed ID
23161513
Organism
Homo sapiens
Experiment description
Proteomic analysis of exosomes from mutant KRAS colon cancer cells identifies intercellular transfer of mutant KRAS.
Authors
Demory Beckler M, Higginbotham JN, Franklin JL, Ham AJ, Halvey PJ, Imasuen IE, Whitwell C, Li M, Liebler DC, Coffey RJ.
Journal name
Mol Cell Proteomics
Publication year
2012
Sample
Colorectal cancer cells
Sample name
Dks-8
Isolation/purification methods
Differential centrifugation Filtration
Flotation density
-
Molecules identified in the study
Protein
Methods used in the study
Mass spectrometry
7
Experiment ID
209
MISEV standards
✘
EV Biophysical techniques
✔
TSG101|HSP70|FLOT1
EV Enriched markers
✔
VDAC
EV Negative markers
✔
NTA
EV Particle analysis
Identified molecule
protein
Identification method
Mass spectrometry
PubMed ID
23161513
Organism
Homo sapiens
Experiment description
Proteomic analysis of exosomes from mutant KRAS colon cancer cells identifies intercellular transfer of mutant KRAS.
Authors
Demory Beckler M, Higginbotham JN, Franklin JL, Ham AJ, Halvey PJ, Imasuen IE, Whitwell C, Li M, Liebler DC, Coffey RJ.
Journal name
Mol Cell Proteomics
Publication year
2012
Sample
Colorectal cancer cells
Sample name
DLD-1
Isolation/purification methods
Differential centrifugation Filtration
Flotation density
-
Molecules identified in the study
Protein
Methods used in the study
Mass spectrometry
8
Experiment ID
1203
MISEV standards
✔
EM
EV Biophysical techniques
✔
SDCBP|FLOT1|CD9|CD81|CD63|EPCAM|GAPDH|LAMP1|TFRC|CD151|CD82|LAMP2|RAB35|TSG101|FLOT2|RAB5B|ICAM1|RAB5A
EV Enriched markers
✘
EV Negative markers
✘
EV Particle analysis
Identified molecule
protein
Identification method
Mass spectrometry
PubMed ID
34887515
Organism
Homo sapiens
Experiment description
Supermeres are functional extracellular nanoparticles replete with disease biomarkers and therapeutic targets
Authors
Zhang Q, Jeppesen DK, Higginbotham JN, Graves-Deal R, Trinh VQ, Ramirez MA, Sohn Y, Neininger AC, Taneja N, McKinley ET, Niitsu H, Cao Z, Evans R, Glass SE, Ray KC, Fissell WH, Hill S, Rose KL, Huh WJ, Washington MK, Ayers GD, Burnette DT, Sharma S, Rome LH, Franklin JL, Lee YA, Liu Q, Coffey RJ.
Journal name
Nat Cell Biol
Publication year
2021
Sample
Colorectal cancer cells
Sample name
DiFi
Isolation/purification methods
Differential centrifugation Filtration Centrifugal ultrafiltration Ultracentrifugation OptiPrep density gradient centrifugation
Flotation density
-
Molecules identified in the study
Protein miRNA
Methods used in the study
Western blotting Mass spectrometry RNA sequencing
9
Experiment ID
363
MISEV standards
✘
EV Biophysical techniques
✔
TSG101|GAPDH|AQP1|CD151|CD81|CD82|CD9|EPCAM|FLOT1|FLOT2|ICAM1|ITGA2B|LAMP2|RAB35|RAB5A|RAB5B|SDCBP|TFRC|UCHL1
EV Enriched markers
✔
DCLK1
EV Negative markers
✔
NTA
EV Particle analysis
Identified molecule
Protein
Identification method
Mass spectrometry
PubMed ID
33991177
Organism
Homo sapiens
Experiment description
Cancer stem cell marker DCLK1 reprograms small extracellular vesicles toward migratory phenotype in gastric cancer cells
Authors
Carli ALE, Afshar-Sterle S, Rai A, Fang H, O'Keefe R, Tse J, Ferguson FM, Gray NS, Ernst M, Greening DW, Buchert M.
Journal name
Proteomics
Publication year
2021
Sample
Gastric cancer cells
Sample name
MKN1 - 100K pellet
Isolation/purification methods
Differential centrifugation Ultracentrifugation
Flotation density
-
Molecules identified in the study
Protein
Methods used in the study
Western blotting Mass spectrometry
10
Experiment ID
364
MISEV standards
✘
EV Biophysical techniques
✔
TSG101|GAPDH|AQP1|CD151|CD81|CD82|CD9|EPCAM|FLOT1|FLOT2|ICAM1|ITGA2B|LAMP2|RAB35|RAB5A|RAB5B|SDCBP|TFRC|UCHL1
EV Enriched markers
✔
DCLK1
EV Negative markers
✔
NTA
EV Particle analysis
Identified molecule
Protein
Identification method
Mass spectrometry
PubMed ID
33991177
Organism
Homo sapiens
Experiment description
Cancer stem cell marker DCLK1 reprograms small extracellular vesicles toward migratory phenotype in gastric cancer cells
Authors
Carli ALE, Afshar-Sterle S, Rai A, Fang H, O'Keefe R, Tse J, Ferguson FM, Gray NS, Ernst M, Greening DW, Buchert M.
Journal name
Proteomics
Publication year
2021
Sample
Gastric cancer cells
Sample name
MKN1 - 100K pellet
Isolation/purification methods
Differential centrifugation Ultracentrifugation
Flotation density
-
Molecules identified in the study
Protein
Methods used in the study
Western blotting Mass spectrometry
11
Experiment ID
365
MISEV standards
✘
EV Biophysical techniques
✔
TSG101|GAPDH|AQP1|CD151|CD81|CD82|CD9|EPCAM|FLOT1|FLOT2|ICAM1|ITGA2B|LAMP2|RAB35|RAB5A|RAB5B|SDCBP|TFRC|UCHL1
EV Enriched markers
✔
DCLK1
EV Negative markers
✔
NTA
EV Particle analysis
Identified molecule
Protein
Identification method
Mass spectrometry
PubMed ID
33991177
Organism
Homo sapiens
Experiment description
Cancer stem cell marker DCLK1 reprograms small extracellular vesicles toward migratory phenotype in gastric cancer cells
Authors
Carli ALE, Afshar-Sterle S, Rai A, Fang H, O'Keefe R, Tse J, Ferguson FM, Gray NS, Ernst M, Greening DW, Buchert M.
Journal name
Proteomics
Publication year
2021
Sample
Gastric cancer cells
Sample name
MKN1 - 100K pellet
Isolation/purification methods
Differential centrifugation Ultracentrifugation
Flotation density
-
Molecules identified in the study
Protein
Methods used in the study
Western blotting Mass spectrometry
12
Experiment ID
235
MISEV standards
✔
EM
EV Biophysical techniques
✔
TSG101|Alix|HSC70|GAPDH
EV Enriched markers
✔
HSP90B1
EV Negative markers
✔
qNano
EV Particle analysis
Identified molecule
protein
Identification method
Mass spectrometry
PubMed ID
26054723
Organism
Homo sapiens
Experiment description
Hepatocellular carcinoma-derived exosomes promote motility of immortalized hepatocyte through transfer of oncogenic proteins and RNAs
Authors
He M, Qin H, Poon TC, Sze SC, Ding X, Co NN, Ngai SM, Chan TF, Wong N
Journal name
Carcinogenesis
Publication year
2015
Sample
Hepatocellular carcinoma cells
Sample name
HKCI-8
Isolation/purification methods
Differential centrifugation Filtration Ultracentrifugation Sucrose density gradient
Flotation density
1.13-1.19 g/mL
Molecules identified in the study
Protein RNA
Methods used in the study
Western blotting Mass spectrometry RT-PCR RNA Sequencing
13
Experiment ID
126
MISEV standards
✘
EV Biophysical techniques
✔
GAPDH
EV Enriched markers
✘
EV Negative markers
✘
EV Particle analysis
Identified molecule
protein
Identification method
Mass spectrometry [LTQ-FT Ultra]
PubMed ID
Unpublished / Not applicable
Organism
Homo sapiens
Experiment description
Mesenchymal Stem Cell Exosomes: The Future MSC-based Therapy?
Authors
Ruenn Chai Lai, Ronne Wee Yeh Yeo, Soon Sim Tan, Bin Zhang, Yijun Yin, Newman Siu Kwan Sze, Andre Choo, and Sai Kiang Lim
Journal name
Mesenchymal Stem Cell Therapy
Publication year
2011
Sample
Mesenchymal stem cells
Sample name
huES9.E1
Isolation/purification methods
HPLC
Flotation density
-
Molecules identified in the study
Protein
Methods used in the study
Western blotting Antobody array Mass spectrometry
14
Experiment ID
488
MISEV standards
✔
EM
EV Biophysical techniques
✔
CD9|CD81|CD63|GAPDH|SDCBP|LAMP1|TFRC|UCHL1|FLOT2|LAMP2|FLOT1|ICAM1|RAB5B|CD151|RAB35|TSG101|RAB5A|CD82
EV Enriched markers
✔
CANX
EV Negative markers
✔
NTA
EV Particle analysis
Identified molecule
Protein
Identification method
Mass spectrometry
PubMed ID
36408942
Organism
Homo sapiens
Experiment description
Quantitative proteomic analysis of exosomes from umbilical cord mesenchymal stem cells and rat bone marrow stem cells
Authors
Xu X, Yin F, Guo M, Gan G, Lin G, Wen C, Wang J, Song P, Wang J, Qi ZQ, Zhong CQ.
Journal name
Proteomics
Publication year
2023
Sample
Mesenchymal stem cells
Sample name
UCMSC
Isolation/purification methods
Differential centrifugation Filtration Ultracentrifugation
Flotation density
-
Molecules identified in the study
Protein
Methods used in the study
Western blotting Mass spectometry
15
Experiment ID
211
MISEV standards
✔
EM
EV Biophysical techniques
✔
TSG101|Alix|EpCAM|TFRC
EV Enriched markers
✔
cytochrome c|GOLGA2
EV Negative markers
✘
EV Particle analysis
Identified molecule
protein
Identification method
Mass spectrometry
PubMed ID
23333927
Organism
Homo sapiens
Experiment description
Characterization and proteomic analysis of ovarian cancer-derived exosomes.
Authors
Liang B, Peng P, Chen S, Li L, Zhang M, Cao D, Yang J, Li H, Gui T, Li X, Shen K.
Journal name
J Proteomics
Publication year
2013
Sample
Ovarian cancer cells
Sample name
IGROV1
Isolation/purification methods
Differential centrifugation Ultracentrifugation Sucrose density gradient
Flotation density
1.09-1.15 g/mL
Molecules identified in the study
Protein
Methods used in the study
Mass spectrometry
16
Experiment ID
212
MISEV standards
✔
CEM
EV Biophysical techniques
✔
TSG101|Alix|EpCAM|TFRC
EV Enriched markers
✔
Cytochrome C|GOLGA2
EV Negative markers
✘
EV Particle analysis
Identified molecule
protein
Identification method
Mass spectrometry
PubMed ID
23333927
Organism
Homo sapiens
Experiment description
Characterization and proteomic analysis of ovarian cancer-derived exosomes.
Authors
Liang B, Peng P, Chen S, Li L, Zhang M, Cao D, Yang J, Li H, Gui T, Li X, Shen K.
Journal name
J Proteomics
Publication year
2013
Sample
Ovarian cancer cells
Sample name
OVCAR-3
Isolation/purification methods
Differential centrifugation Ultracentrifugation Sucrose density gradient
Flotation density
1.09-1.15 g/mL
Molecules identified in the study
Protein
Methods used in the study
Mass spectrometry
17
Experiment ID
622
MISEV standards
✔
EM
EV Biophysical techniques
EV Enriched markers
✔
GOLGA2|CYC1
EV Negative markers
✔
NTA
EV Particle analysis
Identified molecule
Protein
Identification method
Mass spectrometry
PubMed ID
29339034
Organism
Homo sapiens
Experiment description
Extracellular matrix proteins and carcinoembryonic antigen-related cell adhesion molecules characterize pancreatic duct fluid exosomes in patients with pancreatic cancer
Authors
Zheng J, Hernandez JM, Doussot A, Bojmar L, Zambirinis CP, Costa-Silva B, van Beek EJAH, Mark MT, Molina H, Askan G, Basturk O, Gonen M, Kingham TP, Allen PJ, D'Angelica MI, DeMatteo RP, Lyden D, Jarnagin WR.
Journal name
HPB
Publication year
2018
Sample
Pancreatic duct fluid
Sample name
From patients with Pancreatic ductal adenocarcinoma
Isolation/purification methods
Differential centrifugation Ultracentrifugation
Flotation density
-
Molecules identified in the study
Protein
Methods used in the study
Mass spectrometry
18
Experiment ID
623
MISEV standards
✘
EV Biophysical techniques
EV Enriched markers
✔
GOLGA2|CYC1
EV Negative markers
✔
NTA
EV Particle analysis
Identified molecule
Protein
Identification method
Mass spectrometry
PubMed ID
29339034
Organism
Homo sapiens
Experiment description
Extracellular matrix proteins and carcinoembryonic antigen-related cell adhesion molecules characterize pancreatic duct fluid exosomes in patients with pancreatic cancer
Authors
Zheng J, Hernandez JM, Doussot A, Bojmar L, Zambirinis CP, Costa-Silva B, van Beek EJAH, Mark MT, Molina H, Askan G, Basturk O, Gonen M, Kingham TP, Allen PJ, D'Angelica MI, DeMatteo RP, Lyden D, Jarnagin WR.
Journal name
HPB
Publication year
2018
Sample
Pancreatic duct fluid
Sample name
From patients with IPMN with microinvasive cancer (IPMNca)
Isolation/purification methods
Differential centrifugation Ultracentrifugation
Flotation density
-
Molecules identified in the study
Protein
Methods used in the study
Mass spectrometry
19
Experiment ID
275
MISEV standards
✔
EM
EV Biophysical techniques
✔
TSG101|Alix|RAB5A|CD9|CD82|CD63|CD81
EV Enriched markers
✔
AIF
EV Negative markers
✔
NTA
EV Particle analysis
Identified molecule
protein
Identification method
Mass spectrometry
PubMed ID
25844599
Organism
Homo sapiens
Experiment description
Molecular profiling of prostate cancer derived exosomes may reveal a predictive signature for response to docetaxel.
Authors
Kharaziha P, Chioureas D, Rutishauser D, Baltatzis G, Lennartsson L, Fonseca P, Azimi A, Hultenby K, Zubarev R, Ullen A, Yachnin J, Nilsson S, Panaretakis T.
Journal name
Oncotarget
Publication year
2015
Sample
Prostate cancer cells
Sample name
DU145 - Docetaxel sensitive
Isolation/purification methods
Filtration Ultracentrifugation Sucrose density gradient
Flotation density
1.12-1.19 g/mL
Molecules identified in the study
Protein
Methods used in the study
Mass spectrometry/Flow cytometry/Western blotting
20
Experiment ID
274
MISEV standards
✔
EM
EV Biophysical techniques
✔
TSG101|Alix|RAB5A|CD9|CD82|CD63|CD81
EV Enriched markers
✔
AIF
EV Negative markers
✔
NTA
EV Particle analysis
Identified molecule
protein
Identification method
Mass spectrometry
PubMed ID
25844599
Organism
Homo sapiens
Experiment description
Molecular profiling of prostate cancer derived exosomes may reveal a predictive signature for response to docetaxel.
Authors
Kharaziha P, Chioureas D, Rutishauser D, Baltatzis G, Lennartsson L, Fonseca P, Azimi A, Hultenby K, Zubarev R, Ullen A, Yachnin J, Nilsson S, Panaretakis T.
Journal name
Oncotarget
Publication year
2015
Sample
Prostate cancer cells
Sample name
DU145 - Docetaxel resistant
Isolation/purification methods
Filtration Ultracentrifugation Sucrose density gradient
Flotation density
1.13-1.18 g/mL
Molecules identified in the study
Protein
Methods used in the study
Mass spectrometry Flow cytometry Western blotting
21
Experiment ID
834
MISEV standards
✔
EM
EV Biophysical techniques
✔
LAMP2|CD63|FLOT1|ITGA2B|ICAM1|CD9|CD151|TFRC|RAB5A|GAPDH|AQP1|TSG101
EV Enriched markers
✔
CANX
EV Negative markers
✔
NTA
EV Particle analysis
Identified molecule
Protein
Identification method
Mass spectrometry
PubMed ID
35333565
Organism
Homo sapiens
Experiment description
LAMP2A regulates the loading of proteins into exosomes
Authors
Ferreira JV, da Rosa Soares A, Ramalho J, Máximo Carvalho C, Cardoso MH, Pintado P, Carvalho AS, Beck HC, Matthiesen R, Zuzarte M, Girão H, van Niel G, Pereira P
Journal name
Sci Adv
Publication year
2022
Sample
Retinal pigment epithelial cells
Sample name
ARPE-19
Isolation/purification methods
Differential centrifugation Filtration Ultracentrifugation
Flotation density
-
Molecules identified in the study
Protein
Methods used in the study
Western blotting Mass spectrometry
22
Experiment ID
835
MISEV standards
✔
EM
EV Biophysical techniques
✔
CD63|FLOT1|ITGA2B|ICAM1|CD9|CD151|TFRC|RAB5A|GAPDH|AQP1|TSG101
EV Enriched markers
✔
CANX
EV Negative markers
✔
NTA
EV Particle analysis
Identified molecule
Protein
Identification method
Mass spectrometry
PubMed ID
35333565
Organism
Homo sapiens
Experiment description
LAMP2A regulates the loading of proteins into exosomes
Authors
Ferreira JV, da Rosa Soares A, Ramalho J, Máximo Carvalho C, Cardoso MH, Pintado P, Carvalho AS, Beck HC, Matthiesen R, Zuzarte M, Girão H, van Niel G, Pereira P
Journal name
Sci Adv
Publication year
2022
Sample
Retinal pigment epithelial cells
Sample name
ARPE-19
Isolation/purification methods
Differential centrifugation Filtration Ultracentrifugation
Flotation density
-
Molecules identified in the study
Protein
Methods used in the study
Western blotting Mass spectrometry
Protein-protein interactions for ADAM9
Protein Interactor
ExoCarta ID
Identification method
PubMed
Species
1
SNX9
51429
Two-hybrid
Homo sapiens
Reconstituted Complex
Homo sapiens
Affinity Capture-Western
Homo sapiens
2
PRKCD
5580
Affinity Capture-Western
Homo sapiens
3
MAD2L2
10459
Two-hybrid
Homo sapiens
Reconstituted Complex
Homo sapiens
4
SH3GL2
Two-hybrid
Homo sapiens
Reconstituted Complex
Homo sapiens
Affinity Capture-Western
Homo sapiens
View the network
image/svg+xml
Pathways in which ADAM9 is involved