Figure 1
Non-muscle Myosin IIA is the governor of EtOH- and Sar1a-KD-induced Golgi fragmentation.
(a) Immunostaining of giantin in VA-13 cells (HepG2 cells expressing ADH). First row: no treatment (Ctrl), EtOH treatment (35 mM for 72 h) and EtOH treatment followed by 25 μM blebbistatin at 48 h; second row: control siRNA and 35 mM EtOH for 72 h plus control or MYH9 siRNAs; third row: MYH9 siRNAs, SAR1A siRNAs or SAR1A+MYH9 siRNAs. The right panel shows high magnifications of the highlighted area (white boxes). Nuclei were counterstained with DAPI (blue). (b) NMIIA Western blot of the lysates of VA-13 cells treated with 35 mM EtOH for 72 h; β-actin was a loading control. (c) Quantitative real-time PCR analysis of the mRNA of NMIIA in control VA-13 cells and treated with 35 mM EtOH for 72 h. Data were presented as a mean from the three independent experiments and expressed as the fold relative to that (100%, 1 fold) of GAPDH. (d) NMIIA Western blot of the lysates of VA-13 cells treated with scramble or MYH9 siRNAs; β-actin was a loading control. (e) NMIIA and Sar1a Western blot of the lysates of VA-13 cells treated with scramble or SAR1A plus NMIIA siRNAs; β-actin was a loading control. (f) Quantification of the fragmented Golgi in cells treated as described in the (a) n = 60 cells from three independent experiments.
