Fig. 6: DDX3X promotes FAM134B transcription.

A Western blot detected FAM134B protein level in Huh7 and MHCC97-H cells stably expressing shNC or shDDX3X. B MHCC97-H cells were transfected with vector, Flag-WT DDX3X, Flag-mutant DDX3X at the same concentration. Forty-eight hours later, cells were lysed with RIPA, followed by western blot analysis. C Huh7 cells stably expressing either shNC or shDDX3X were treated with CHX (20 μM) for 0, 2, 4, 8 h. The degradation kinetics of FAM134B was monitored over time by western blot. Data are mean ± SEM from three independent experiments, ns: not significant. D MHCC97-H cells stably expressing either shNC or shDDX3X were treated with CHX (20 μM) for 0, 2, 4, 8 h. The degradation kinetics of FAM134B was monitored over time by western blot. Data are mean ± SEM from three independent experiments, ns: not significant. E MHCC97-H and Huh7 cells were transfected with siNC or siDDX3X. Twenty-four hours later, total RNA was isolated using TRIzol reagent. RT-qPCR was then employed to assess the mRNA levels of DDX3X and FAM134B. Data are mean ± SEM from three independent experiments, ***P < 0.001. F, G Luciferase reporter assay detected the effect of DDX3X on FAM134B promoter activity. Data are mean ± SEM from three independent experiments, **P < 0.01. H The representative IHC images of DDX3X protein expression in HCC tissues. Scale bar, 40 μm.