Fig. 3

RanBPM requires HDAC6 for association with α-tubulin. a Control or RanBPM shRNA Hela cells were fixed and incubated with antibodies against HDAC6 (green) and α-tubulin (red). Colocalization of HDAC6 and α-tubulin was analyzed using Imaris software. Top, representative images where white signal represents the top 2% of HDAC6 and α-tubulin colocalization. Bottom left, quantification of the top 2% of pixels representing HDAC6 colocalized with α-tubulin. Data are representative of a minimum of 60 cells from three experiments. Error bars represent SEM. P < 0.05 (*). Bottom right, western blot analysis of extracts from Hela control and RanBPM shRNA cells, showing RanBPM and HDAC6 expression with respect to vinculin loading control. b Wildtype (WT) or HDAC6 knockout (KO) MEFs were fixed and incubated with antibodies against RanBPM (green) and α-tubulin (red). Colocalization of RanBPM and α-tubulin was analyzed using Imaris software. Top, representative images where white signal represents the top 2% of RanBPM and α-tubulin colocalization. Bottom left, quantification of the top 2% of pixels representing RanBPM colocalized with α-tubulin. Data are representative of a minimum of 60 cells from three experiments. Error bars represent SEM. P < 0.05 (*). Bottom right, western blot analysis of extracts from WT and HDAC6 KO MEFs, showing HDAC6 and RanBPM expression with respect to β-actin loading control