After 6 h, U0126- and IWR-1-treated cells were labeled with EdU (10 nM) and incubated for 1 h. The EdU staining steps were carried out according to the EdU manufacturers instruction manual. Nuclei were subsequently stained with Hoechst 33,342, and EdU-positive cell percentages are shown on the right. HepG2 cells were transfected with pCAGEN-Flag-V (to overexpress the V protein). After 24 h ( A ), U0126 ( B ) or IWR-1 ( C ) was added to cells, which were incubated for another 6 h, followed by labeling with EdU (10 nM) and incubation for 1 h. Then, the cells were stained with rabbit anti-Flag antibody and goat anti-rabbit IgG Alexa Fluor 488 (green) secondary antibody. For EdU staining, the steps were carried out according to the EdU manufacturers instructions. Nuclei were subsequently stained with Hoechst 33,342. Images were captured using a Leica fluorescence microscope (400 x), bar = 50 m
