Fig. 3: RAB10 dephosphorylation is required for its removal from STm invasion sites.

a and b Representative images (a) and quantifications (b) of phospho-RAB10 (T73) signal measured by western blot in WT Henle 407 cells infected with WT STm for indicated time. In this and following panels with the quantification of RAB10 phosphorylation by western blot, the relative phospho-RAB10 (T73) signal was calculated by comparison with the respective total RAB10 signal. c and d, Representative images (c) and quantifications (d) of PM-RAB10 retention at STm invasion sites at 30 min p.i. RAB10 KO Henle 407 cells were transfected with WT, T73E or T73A myc-PM-RAB10, and then infected with WT STm. Cells were fixed at 30 min p.i. and stained for myc-tag. In this and the following panels, invasion sites at 30 min p.i. were identified by staining of extracellular STm before permeabilization. Line plot profile follows the white arrow in the ‘Merge’ channel in c. e and f, Representative images (e) and quantifications (f) of GFP-RAB10 retention at STm invasion sites at 30 min p.i. RAB10 KO Henle 407 cells were transfected with GFP-RAB10 WT, Q68L or T23N construct, and then infected with WT STm. Cells were fixed and imaged at 30 min p.i. Data shown are means ± S.D. for three independent experiments. At least 100 invasion sites (d and f) for each condition in each experiment were scored for the retention of RAB10. P values were calculated using one-way ANOVA. Scale bars, 10 μm. Source data are provided as a Source Data file.