Fig. 4

(A-C) Cycloheximide chase assays. Following Wnt pathway inhibitor XAV939 (A) and IWR-1 (B) treatment for 24 h or shRNA knockdown (C) for 48 h, HuH7 cells were stimulated with IFN-γ for 24 h. We then treated the cells with 20 µg/ml cycloheximide (CHX) for 0, 2, 4, and 8 h prior to cell harvest. Western blotting was utilized to detect the protein amount. PD-L1 level is reported as fold changes relative to cells treated with vehicle only (A-B) or scramble shRNAs (C) and without CHX. (D-G) Western blotting demonstrating changes of eIF-4E and 4E-BP1 levels along with PD-L1 expression after Wnt pathway inhibitor treatment. HuH7 (D-E) and Hep3B (F-G) cells were treated with XAV939 (D and F) or IWR-1 (E and G) for 24 h before 25 ng/mL IFN-γ was added. The cells were harvested after another 24 h