Fig. 1

EZH2 negatively regulates the IFNγ-induced PD-L1 expression. a Representative IHC staining of EZH2 in HCC tissues. The black arrows indicate the expression of EZH2 on stroma cells, and the red arrows indicate the expression of EZH2 on TCs. b Representative pictures of multiple immunofluorescence staining showing the expression of EZH2 (green) and CD68 (red) in HCC. Scale bar, 50 μm. The white arrows indicate Mo/Mφs, and the five-pointed stars indicate TCs. c Immunoblotting analysis showing the expression of IFNγ-induced PD-L1 in hepatoma cells and monocytes. d Hep3B cells were transfected with negative control (NC) or different EZH2-targeted siRNAs for 48 h, and then treated with IFNγ for 24 h. Immunoblotting analyses were performed to detect the expression of EZH2 and PD-L1. β-actin was used as a loading control. e qPCR analysis showing that downregulation of EZH2 promoted the mRNA expression of IFNγ-induced PD-L1 in PLC/PRF/5, Huh7, and Hep3B cells. f FACS staining showing that downregulation of EZH2 promoted the expression of IFNγ-induced PD-L1 in PLC/PRF/5, Huh7, and Hep3B cells. g Downregulation of EZH2 increased the protein level of IFNγ-induced PD-L1 in PLC/PRF/5, Huh7, and Hep3B cells. The indicated hepatoma cells were transfected with EZH2-targeted or NC siRNA for 48 h, and then treated with IFNγ for an additional 24 h. Immunoblotting analysis was performed to detect the protein levels of PD-L1, EZH2, and H3K27me3. GAPDH was used as a loading control