Figure S5 Single color confocal microscopy images from Figure 3. (A) ECFP-Rab7 (blue). (B) LAMP1-EYFP (green). (C) Antibody against M6PR labeled with a Cy5 secondary antibody (red). (TIF) Figure S6 Western blot of BS-C-1 cells stably expressing ECFP-Rab7. BS-C-1 cells and BS-C-1 cells stably expressing ECFP-Rab7 were lysed in a 1% Triton X-100 lysis buffer containing a protease inhibitor (Halt, 78441, Pierce, Rockford, IL) for 30 min at 4uC followed by centrifugation at 14,000 rcf for 20 min at 4uC. BCA analysis was used to determine protein concentration. Lysate was diluted in a Laemmli loading buffer (BP-110R), run on a Tris-glycine SDS gel (456–1094, Bio-Rad, Hercules, CA), and transferred to a PVDF membrane. The membrane was blocked (Near IR Blocking Buffer, MB-070, Rockland Immunochemicals, Gilbertsville, PA) for 1 hr at room temperature. Primary antibodies were incubated overnight at 4uC in blocking buffer and the membrane was washed with TBS- Tween. 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