Localization of endogenous ARHGAP22 in C2C12 cells.

<p>(<b>A</b>) C2C12 cells were fixed and stained with anti-ARHGAP22 (green) or anti-Rab11 (red) antibodies, which was non-treated (control) or preabsorbed with lysates from non-transfected (−) or HA-ARHGAP22-transfected (+) HEK cells. Merged fluorescent images are shown. The cells were also stained with hoechst 33258 for nuclei (blue). Scale bar, 20 µm. Inset shows a magnification image of the boxed region. (<b>B</b>) C2C12 cells were treated with control or <i>ARHGAP22</i> siRNAs for 48 h and serum-starved. The cells were fixed and stained with anti-ARHGAP22 (green) and anti-Rab11 (red) antibodies. Merged fluorescent images are shown. The cells were also stained with hoechst 33258 (blue). Scale bar, 20 µm. (<b>C</b>) C2C12 cells were fixed and stained with anti-ARHGAP22 antibody (green) and antibodies for TGN46 or EEA1 (red). Merged fluorescent images are shown. The cells were also stained with hoechst 33258 (blue). Scale bar, 20 µm. (<b>D</b>) Pearson's Colocalization Coefficient (PCC) was calculated by ImageJ (NIH). The data are expressed as the mean ± s.e.m. (N = 3). Ten cells were analyzed for each experiment. **, <i>p</i><0.01. Statistical significance was determined by Student's <i>t</i>-test.