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. 2016 Apr 27;291(25):12930–12942. doi: 10.1074/jbc.M115.698530

FIGURE 9.

FIGURE 9.

Increased presence of Rab7 in αB-silenced ARPE cells. ARPE cells were immunogold-labeled with anti-Rab5 (early endosome marker) and anti-Rab7 (late endosome marker). A, TEM of four endosomes from control ARPE cells (top panel) shows insignificant Rab5 labeling (12-nm gold particles) and moderately labeled Rab7 (18-nm gold particles). In contrast, there is enhanced immunolabeling of Rab7 (18-nm gold particles) in the endolysosomal compartment with no perceptible change in Rab5 (12-nm gold particles) in αB-silenced (αBshRNA) ARPE cells. Three micrographs of potentially fused vesicles (representing enhanced endolysosomal compartment) are shown. B, quantitation of immunogold labeling by ImageJ (analysis of particles) shows a 5-fold increase in Rab7 labeling but not in the Rab5. C, immunoblots of whole cell extracts for αB, Rab5, Rab7, and HSP70 in native, αB shRNA, transfection control, and scramble control ARPE cells. Note a noticeable increase in Rab7 in αB shRNA lane (white arrow) but no detectable change in Rab5 or HSP70 (run as an internal control). D, quantitation of the immunoblots shown in C. The data in C and D corroborate the TEM data in A and B.