Fig. 6

Knockout of moesin in 786-0 cells inhibits TMIGD1-dependent α-tubulin acetylation and F-actin orientation. A Whole cell lysates from 786-0 cells expressing control sgRNA or moesin-sgRNA (MS-sgRNA) and 786-0 cells co-expressing control sgRNA with TMIGD1 or MS-sgRNA with TMIGD1 were subjected to Western blot via blotting for moesin, acetyl-tubulin, total tubulin, TMIGD1 or GAPDH ***p < 0.001, **p < 0.01. B The same cell lines as panel A were subjected to immunofluorescence staining using anti-acetyl-tubulin antibody. Image magnification, 50 µM. 786-0 cells expressing Ctr. sgRNA, Ctr.sgRNA/TMIGD1 or moesin-sgRNA/TMIGD1 were stained with phalloidin/DAPI and actin fibril orientation/anisotropy was quantified. C EV/786-0 and TMIGD1/786-0 cells were treated with nocodazole (0.5 µg/ml) for various times followed by staining with an antibody against acetylated-α-tubulin. The numbers of cells displaying intact microtubules were manually counted (n = 50 cells per each group of triplicates). EV/786-0 and TMIGD1/786-0 cells were stained with an antibody against acetylated-α-tubulin (green) and phalloidin/actin (red) and pictures of mitotic cells (n = 20 cells per group) were taken under an epifluorescence microscope. ***p < 0.001