Figure 3

Upregulation of PD-L1 in sorafenib-resistant HCC cells. (a) SK-Hep1 and SK-Hep1R cells were treated with or without IFN-γ (15 ng/mL). After 48 h, glucose uptake was measured. Data are shown as the mean of three independent experiments ± SD. **; p < 0.01 (b,c) SK-Hep1 and SK-Hep1R cells were treated with IFN-γ (15 ng/mL). After 48 h treatment, the expression levels of HK2 (b) and CD274 (c) were analyzed by quantitative real-time PCR. The expression levels of target genes were normalized to that of the housekeeping gene GAPDH based on the 2^−ΔΔCt method. Data are shown as the mean of three independent experiments ± SD. **; p < 0.01. (d) SK-Hep1 and SK-Hep1R cells were treated with IFN-γ (15 ng/mL) for 48 h. PD-L1, HK2, and β-actin protein levels were evaluated by WB. The full-length blots are in Supplementary Fig. S7. (e) Densitometry intensity ratios for (d) from replicated WB analyses (n = 3). *; p < 0.05, ***; p < 0.001. (f) SK-Hep1 and SK-Hep1R cells were treated with IFN-γ (15 ng/mL). After 48 h, the cells were treated with the indicated concentration of glucose for 8 h. PD-L1, HK2, and β-actin protein levels were evaluated using WB. The full-length blots are shown in Supplementary Fig. S8. (g) Densitometry intensity ratios for (f) from replicated WB analyses (n = 3). *; p < 0.05, **; p < 0.01, ***; p < 0.001, ****; p < 0.0001.