Extended Data Fig. 3: Phosphorylation promotes Rab10 and Rab18, but not Rab1B, accumulating on the ER.
a, MDA-MB-231 cells co-expressing ER-RFP and GFP-tagged wild-type or mutant Rab1B were analyzed by confocal fluorescent microscopy. Scale bars = 20 μm. b, Colocalization of ER-RFP and GFP-tagged wild-type or mutant Rab1B was quantified in multiple cells, and evaluated by Pearson’s correlation coefficient. WT, n = 22; T72A, n = 19; T72D, n = 27. c, MDA-MB-231 cells co-expressing ER-RFP and GFP-tagged wild-type or mutant Rab10 were analyzed by confocal fluorescent microscopy. Scale bars = 20 μm. d, Colocalization of ER-RFP and GFP-tagged wild-type or mutant Rab10 was quantified in multiple cells, and evaluated by Pearson’s correlation coefficient. WT, n = 15; T73A, n = 15; T73D, n = 21. e, MDA-MB-231 cells co-expressing ER-RFP and GFP-tagged wild-type or mutant Rab18 were analyzed by confocal fluorescent microscopy. Scale bars = 20 μm. f, Colocalization of ER-RFP and GFP-tagged wild-type or mutant Rab18 was quantified in multiple cells, and evaluated by Pearson’s correlation coefficient. WT, n = 16; T72A, n = 17; T72D, n = 16. g, Whole cell lysate and ER extract from MDA-MB-231 cells were analyzed by immunoblotting. Data shown in g are representative of two independent experiments. Data from a-f were examined over three independent experiments. Data from b, d, f were analyzed using one-way Welch’s ANOVA test (P = 0.2830 in b, P < 0.0001 in d and f), and pairwise comparisons were performed using Dunnett’s T3 multiple comparisons test. Data are presented as mean ± s.e.m.
