Fig. 4
Rab32 Promotes Autophagic Degradation of MAM Proteins. A Immunoblot analysis of transfected MCF7 cells with and without 100 nM Bafilomycin A1 incubation for the amounts of MAM proteins. TMX1, VDAC1, FACL4, GPx8 and Ero1α were analyzed. Tubulin acts as a loading control, while FLAG signal indicates transfected Rab32. Densitometry analysis of a minimum n = 3 independent experiments. ***p ≤ 0.0001. B Immunofluorescence analysis of TMX1 incorporation into LC3B-decorated structures. Images and insets show control and Rab32Q85L-transfected cells processed for anti-FLAG (blue), anti-LC3B (red) and anti-TMX1 (green) signals. C Immunoblot analysis of transfected MCF7 cells for the amounts of additional MERC proteins. FACL4, VDAC1, and IP3R3 were analyzed. Tubulin acts as a loading control, while FLAG signal indicates transfected Rab32. n = 3 ***p ≤ 0.0001 **p ≤ 0.01 *p ≤ 0.05. D Immunoblot analysis of transfected wild type MEFs and ATG4b knockout MEFs with and without 100 nM Bafilomycin A1 incubation for the amounts of MAM protein TMX1, and LC3. Tubulin was used as loading control. E Immunoblot and densitometry analysis of MCF7 cells transfected with control siRNA or siRNA against Rab32, analyzed for MAM protein TMX1 and Tubulin after 4 h EBSS incubation. n = 3, *p ≤ 0.05