Figure S5.

Rab27a is required for autophagy cargo receptor secretion in response to lysosome inhibition. (A) WCL from serum-starved HEK293 Ts that stably express shRNAs targeting SNAP29 (shSNAP29¹; shSNAP29²) or control shRNA (shNT) and corresponding to secretion experiments in Fig. 6, E and F, were immunoblotted for the indicated proteins. (B) WCL from serum-starved HEK293Ts that stably express shRNAs targeting VAMP8 (shVAMP8¹; shVAMP8²) or control shRNA (shNT) and corresponding to secretion experiments in Fig. 6, E and F were immunoblotted for the indicated proteins. (C) WCL from serum-starved HEK293Ts that stably express shRNAs targeting Rab27a (Rab27a¹; Rab27a²) or control shRNA (NT) and corresponding to secretion experiments in Fig. 6, E and F, were immunoblotted for the indicated proteins. (D) Representative fluorescence micrographs from cells expressing control shRNA (shNT) or shRNA targeting Rab27a (shRab27a) treated with 20 nM BafA1 and immunostained for endogenous p62 (green) and CD63 (magenta). Scale bar, 10 μm; inset scale bar, 2 μm. (E) Scatter plot of Mander’s coefficients for the co-occurrence of p62 with CD63 in BafA1 treated cells expressing control shRNA (shNT) or Rab27a shRNA (shRab27a) in C. Statistical significance was calculated by an unpaired two-tailed student’s t test (mean ± SEM; shNT, n = 10; shRab27a, n = 10; *, P < 0.05). Source data are available for this figure: SourceData FS5.