Fig. 5: Downregulation of PSD3 resulted in partial loss of ARF6 activation.
From: PSD3 downregulation confers protection against fatty liver disease
Human hepatoma Huh7 cells were transiently transfected with negative control SCR siRNA, PSD3 siRNA or ARF6 siRNA. Forty-eight hours after transfection, the cell lysates were incubated with GGA3 PBD agarose beads that selectively isolate and pull down endogenous active ARF6 (ARF6–GTP). a, After precipitation, active ARF6–GTP was detected by immunoblotting using an anti-ARF6 antibody provided in the kit. Cells transfected with ARF6 siRNA were used as a control. The experiment was performed independently six times with similar results. Representative blot presented. b, Knockdown efficiency with ~60% reduction for PSD3 and ~75% for ARF6 as evaluated by quantitative retro transcription PCR analysed by the 2−ΔΔCt method (n = 6). c, Relative ARF6–GTP (active) calculated as GTP–ARF6/calnexin (CNX) (n = 6). Data are shown as mean ± s.d. of the reported independent experiments. Two-sided P values calculated by Mann–Whitney non-parametric test. RU, relative units.
