Fig 2: SOX2 increased JAK-STAT pathway activity and ISG.RS expression. (A, B) The expression of ISG.RS mRNA assessed by qPCR. MM200 cells were transiently transfected with indicated siRNA for 48 hours. Genes were shown for transcripts that were decreased more than twofold. (C, D) Histogram of the percentage of cleaved caspase-3 positive cells among each group. ME4405 and MM200 cells were transfected with indicated shRNA, treated with or without 1000 IU/mL IFNγ for 24 hours and then cocultured with T cells for 6–8 hours (tumor: T=10:1). (E) Change of the p-JAK1/2, p-STAT1/3 level determined by Western blot. ME4405 and MM200 cells were transfected with indicated siRNA for 24 hours and then treated with or without 1000 IU/mL IFNγ for another 24 hours. (F) Change of PTNP1 and SOCS3 mRNA determined by qPCR. SOX2 knock-out B16/F10 cell clones were treated with or without 1000 IU/mL IFNγ for 24 hours. (G) The transcription of PTNP1 and SOCS3 assessed by luciferase report assay. HEK293T cells were transiently transfected with indicated siRNA. Tumor: T, Tumor cells: T cells. Error bars indicate SEM. One-way ANOVA test. ANOVA, analysis of variance; NS, not significant; SOCS3, suppressor of cytokine signaling; ***p<0.001, **p<0.01, *p<0.05.