Fig. 2 | Signal Transduction and Targeted Therapy

Fig. 2

From: VCP downstream metabolite glycerol-3-phosphate (G3P) inhibits CD8+T cells function in the HCC microenvironment

Fig. 2

VCP inhibits CD8+T cells function via metabolite-mediated mechanism. a OT-1 CD8+T cells co-cultured with shCtrl or shVcp Hepa1-6-OVA cells. b Cytotoxicity assessed by measuring lactate dehydrogenase (LDHA) release from Hepa1-6-OVA cells (n = 3). c The percentage of Annexin v+ CD8+T cells co-cultured with shCtrl or shVcp Hepa1-6-OVA cells was determined by flow cytometry (n = 3). d The percentage of cytokines produced by CD8+T cells co-cultured with tumor cells was measured by flow cytometry (n = 3). e Schematic diagram showing OT-1 CD8+T cells co-cultured with shCtrl or shVcp Hepa1-6-OVA cells using a Transwell system. f The percentage of Annexin v+ CD8+T cells in Transwell system was determined by flow cytometry (n = 3). g The proliferation of CD8+T cells in Transwell system was measured by CFSE assay. h The percentage of cytokines produced by CD8+T cells in Transwell system was measured by flow cytometry (n = 3). i Schematic diagram showing CD8+T cells cultured with conditioned medium (CM) produced by shCtrl or shVcp Hepa1-6-OVA cells. j The proliferation of CD8+T cells cultured with CM produced by shCtrl or shVcp Hepa1-6-OVA cells was measured by CFSE assay. k The percentage of cytokines produced by CD8+T cells cultured with CM produced by shCtrl or shVcp Hepa1-6-OVA cells was measured by flow cytometry (n = 3). l Secreted IFN-γ levels by CD8+T cells cultured with CM produced by shCtrl or shVcp Hepa1-6-OVA cells (n = 3). m Schematic diagram showing CD8+T cells cultured with >/<3 kDa CM produced by shCtrl or shVcp Hepa1-6-OVA cells. n The percentage of cytokines produced by CD8+T cells cultured with >/<3 kDa CM produced by shCtrl or shVcp Hepa1-6-OVA cells was measured by flow cytometry (n = 3). o The proliferation of CD8+T cells cultured with >/<3 kDa CM produced by shCtrl or shVcp Hepa1-6-OVA cells was measured by CCK8 assay (n = 3). Two-way ANOVA. p The proliferation of human CD8+T cells cultured with >/<3 kDa CM produced by shCtrl or shVCP HCCLM3 cells was measured by CFSE assay. q The percentage of cytokines produced by human CD8+T cells cultured with >/<3 kDa CM produced by shCtrl or shVCP HCCLM3 cells was measured by flow cytometry (n = 3). r The expression of activation indicators in human CD8+T cells cultured with >/<3 kDa CM produced by shCtrl or shVCP HCCLM3 cells was determined by flow cytometry (n = 3). Data are presented as mean values ± SD. Statistical significance was determined using two-sided t-tests, *P < 0.05, **P < 0.01, and ***P < 0.001

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