Figure 3. Expression of CA-Lyn increases the numbers of autophagosomes per cell and the levels of LC3B-II protein in nutrient-deprived U87 GBM cells.

U87 GBM cells expressing CA-Lyn, DN-Lyn or the LV control and GFP downstream of the IRES were transduced with RFP-LC3 lentiviral vector then plated and subjected to nutrient deprivation as in Figure 1A. A-B, After 5 days, cells were fixed and stained with DAPI nuclear dye. Representative photomicrographs (Mag X40; Scale bar 10 µm) are shown in (A) with arrows indicating autophagosomes (puncta). The numbers of puncta per cell, counted in at least 25 cells for each construct, are shown in (B) in which data are presented as the mean ± SEM with analysis using one-way ANOVA. C-D, At the indicated time-points after initiation of nutrient deprivation, whole cell lysates were western blotted with the indicated antibodies.