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. 2017 May 26;36(14):2018–2033. doi: 10.15252/embj.201797006

Figure 5. Overexpression of E‐Syt2 and E‐Syt3 induces autophagosome formation.

Figure 5

  1. Western blot analysis of the autophagic flux in cell lysates from control (mock) and GFP‐E‐Syt2 or GFP‐E‐Syt3‐expressing HeLa cells, under complete medium and starvation (1 h EBSS) conditions, without or with Bafilomycin A1 (+BAF. A1).
  2. HeLa cells expressing GFP‐E‐Syt2 or GFP‐E‐Syt3 were immunostained for LC3. Compared to control (mock), transfected cells showed a dramatic increase in LC3 puncta, in both basal (complete medium) and starved (1 h) conditions, as evidenced by counting of LC3 puncta (n = 3; 20 cells per condition).
  3. The increase in LC3 puncta observed in cells overexpressing E‐Syt3GFP (similar results were obtained with E‐Syt2GFP, data not shown) involves mainly peripheral rather than perinuclear cellular regions (n = 3). Arrowheads indicate peripheral LC3 puncta (n = 3; 20–70 cells per condition).
Data information: Means ± s.e.m. are plotted. NS, non‐significant, ***P < 0.001, unpaired two‐tailed t‐test. Scale bars, 10 μm.