HRES-1/Rab4 Promotes the Formation of LC3+ Autophagosomes and the Accumulation of Mitochondria during Autophagy
Figure 2
Confocal microscopy of HeLa cells transfected with expression vector producing LC3 tagged with FP650 (emitting red fluorescence) and different isoforms of HRES-1/Rab4, including wild-type (HRES-1/Rab4), C-terminally truncated form (HRES-1/Rab41–121), dominant-negative form (Rab4S27N), constitutively active (HRES-1/Rab4Q72L) and phosphorylation-resistant form (HRES-1/Rab4S204Q) tagged with eGFP (emitting green fluorescence).
Cells were kept in complete medium (Control), starved in serum-free medium without glutamine (Star), or treated with autophagy-modifying agents, bafilomycin A1 (Baf) and rapamycin (Rapa) for 4 hours. Starved and rapamycin treated cells were also treated with Baf. Images are representative of 6–40 cells analyzed during three independent experiments. Image inserts bracketed by broken lines correspond to LC3 vesicles magnified from the original image. Individual and composite color channels are shown for each experimental condition.