Figure 3. Activation of autophagy by combined ER stress and cell death signals. (A–D′) Whole-mount adult retina from flies expressing GFP-LC3 in PRNs. GFP-LC3 is in green and phalloidin labels PRN rhabdomeres in red. (A′–D′) Higher magnification view of GFP staining in the area surrounded by a white rectangle in (A–D). Scale bar ; 10 µm. (E) Quantification of the number of GFP-LC3 dots per PRN. (F) western blot with anti-GFP antibody showing the conversion of GFP-LC3-I to GFP-LC3-II in Drosophila retina. The western blots shown are representative of three independent experiments. (G–J′′) Substantia nigra sections after Tm and 6-OHDA injections in mice. (G–J) Anti-TH in red. (G′-J′) Anti-LC3 in green. (G′′–J′′) The merge shows TH, LC3 and DAPI (blue) for nuclei. (K) The graph shows the percentage of TH positive cells with LC3 punctates. Between 220 and 250 neurons from each of 3 different mice were assessed. Scale bar 20 µm. (L) western blot and quantification showing the conversion of LC3-I to LC3-II in SH-SY5Y cells treated or not treated with Tm and 6-OHDA in the presence of bafilomycin A1. Cells under starvation are used as a positive control. The western blots shown are representative of three independent experiments. *p ≤ 0,05, **p ≤ 0.01 in Student’s t-test.