Rac1 inhibitor suppresses microraft organization during myogenic differentiation. (A) GGS25 cells were cultured for 24 h and then further cultured in pmDM supplemented with 0.1% DMSO or NSC23766 (10–100 μM) for up to 36 h. Differentiated cells were detected by immunostaining with anti-MyHC antibody (filled columns). Fusion indexes were calculated as percentages of nucleus numbers in multinucleated cells (open columns). Averages and standard deviations (n = 3) are shown. (B-E) GGS25 cells were cultured in pmGM (B), pmDM with 0.1% DMSO (C and E), or pmDM with NSC23766 (100 μM) for up to 37 h (D and E). GFP-GPI was sequentially observed at 1 min intervals under epifluorescence microscopy by time-lapse recording. Pictures were taken after 24 h of differentiation cultures (C and D) or growing culture (B). Scale bar, 25 μm. (E) Percentage of cells that showed membrane ruffles during 1 h following the indicated time in pmDM supplemented with DMSO (open columns) or NSC23766 (filled columns). Averages and standard deviations (n = 3) are shown and analyzed using Student’s t-test. *,**P < 0.05.
Mukai and Hashimoto BMC Cell Biology 2013 14:37 doi:10.1186/1471-2121-14-37