To assess the level of cell death, WT MEFs were incubated with 5 nM Bafilomycin A1 (Millipore-sigma) for 48 h or Rab7−/− MEFs expressing control or Rab27 shRNA lentiviruses were treated with DMSO or 50 nM Bafilomycin A1 or Oligomycin + Antimycin A (10 μM + 10 μM) (Sigma-Aldrich) for 24 h. To assess viability after treatments, cells were incubated with Yo-Pro1 (1:1000; Life Technologies) and Hoechst 33342 (1:1000; Invitrogen) for 15 min at 37 °C before imaging. Percent cell death was determined after treatments by dividing the number of Yo-Pro1-positive cells by total number of Hoechst 33342-positive cells as described58 (link).
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