Fig. 3: RAB10 acts locally to generate PM-associated membrane reservoirs and regulate PM biophysical properties.

a 3D rendering of RAB10⁺ PM reservoirs and the PM from a large volume FIB-SEM dataset. b Representative image of a single slice from fluorescence images correlated to the same cell imaged by FIB-SEM. The full correlated dataset is in Supplementary Movie 2. c, d Representative images (c) and quantifications (d) of deep PM reservoirs (as analyzed for the presence of at least one CellMask⁺ tubular structure >10 µm in length) in WT, RAB10 KO or RAB10 KO Henle 407 cells that were transfected with indicated plasmid. GFP-control vectors were co-transfected to indicate transfected cells (marked with green asterisks). n = 3 Independent experiments with 100 cells examined in each experiment. e Atomic force microscopy measurements of basal membrane tension of WT and RAB10 KO Henle 407 cells. n = 3 Independent experiments with ten cells examined in each experiment. f, g Representative images (f) and quantifications (g) of the cell area of WT, RAB10 KO or RAB10 KO Henle 407 cells that were transfected with indicated plasmid. GFP-control vectors were co-transfected to indicate transfected cells and used to quantify cell area. n = 3 Independent experiments with 25 cells examined in each experiment. Data shown are means ± SD. P value was calculated using d, g one-way analysis of variance (ANOVA) and e two-tailed unpaired t test. Scale bars, b 2 μm and c, f 10 μm. Source data are provided as a Source Data file.