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. 2022 Sep 16;298(10):102499. doi: 10.1016/j.jbc.2022.102499

Figure 2.

Figure 2

In vitro extraction assay of Rab10 and Rab29 by GDI1/2. Membrane fractions of HEK293A cells were incubated in the presence of the indicated concentration of recombinant 6His-SUMO1-GDI1 or 6His-SUMO1-GDI2, and the assay mixtures were fractionated into the extracted and not-extracted fractions by ultracentrifugation. Each experiment was carried out side-by-side with the 6His-SUMO1-empty control protein to exclude the effect of the 6His-SUMO1 tag. A and C, representative immunoblots of three independent experiments with the indicated antibodies are shown. B and D, the band intensities of Rab10 and Rab29 were quantified, and the ratio of extracted Rab to total Rab (extracted + not-extracted) was calculated. Rab10 extracted by empty vector and GDI are shown in open and closed circles, respectively. Rab29 extracted by empty vector and GDI are shown in open and closed squares, respectively. The symbols and the error bars represent the mean values and their standard errors, respectively. ∗∗p < 0.01, ∗∗∗p < 0.001. The p-values and statistical tests used are summarized in Table S1. GDI, GDP-dissociation inhibitor.