Fig. 3

CTNNB1 knockdown experiments. Approximately 16 h after seeding, CTNNB1-specific shRNAs or scramble shRNAs were added to HuH7 (A, C, E) and Hep3B (B, D, F) cells. After 48 h, IFN-γ 25 ng/mL was added. The cells were harvested after 24 h. (A-B) Western blotting demonstrating the influence of CTNNB1 knockdown on PD-L1 expression. (C-D) The harvested cells were stained with anti-PD-L1 antibodies or isotype controls. Flow cytometry was used to examine membranous PD-L1 expression. Expression is reported as fold changes relative to cells treated with scramble shRNAs (^*p < 0.05). (E-F)CD274 mRNA expression was analyzed using RT-qPCR with GAPDH as the internal control. Expression is reported as fold changes relative to cells treated with the shRNAs (^*p < 0.05)