Fig. 4: RAB10 effectors contribute to the formation of RAB10⁺ membrane reservoirs.

a The ProHits-viz web tool was used to generate the dot plot view of chosen known and novel RAB10 interactors from the BioID profiling, displaying prey abundance across baits and prey confidence. b Quantifications of RAB10⁺ membrane reservoirs in Henle 407 cells with siRNA knockdown of MICAL-L1 or EHBP1. WT Henle 407 cells were transfected with the indicated siRNA and then transfected with GFP-RAB10 24 h later. n = 4–6 Independent experiments with at least 100 cells examined in each experiment. c Quantifications of BAR domain protein positive tubules in Henle 407 cells transfected with indicated plasmid. n = 3 Independent experiments with 100 cells examined in each experiment. d Representative images of WT Henle 407 cells co-transfected with GFP-RAB10 and PACSIN2-mCherry or PACSIN3-mCherry. Insets were chosen to depict the effectors’ colocalizations on RAB10⁺ membrane reservoirs. MCC values for mCherry-PACSIN3 and GFP-RAB10 are M1 = 0.42 ± 0.08 and M2 = 0.61 ± 0.08. For mCherry-PACSIN2 and GFP-RAB10, M1 = 0.39 ± 0.09 and M2 = 0.42 ± 0.11. e Line plot profiles of the white arrows in the insets in (d). f Quantifications of RAB10⁺ membrane reservoirs in Henle 407 cells with siRNA knockdown of PACSIN2 or PACSIN3. WT Henle 407 cells were transfected with the indicated siRNA and then transfected with GFP-RAB10 24 h later. n = 3 Independent experiments with 100 cells examined in each experiment. Data shown are means ± SD. P value was calculated using (b, f) one-way ANOVA. Scale bars, 10 μm. Source data are provided as a Source Data file.