Fig. 2

Induction and role of autophagy in the response of A549 cells to chemotherapeutics. Cells were treated with chemotherapeutics for 48 h with doses cited in the text, followed by analysis of autophagy markers. a Representative images of LC3 immunocytochemistry. b Percentage of LC3-positive cells. Cells with at least 5 green dots were considered to be positive. U87 glioma cells treated with Rapamycin 100 nM for 24 h were used as a positive control. c Representative plots of Acridine orange (AO) staining. Green (BL1) x red (BL3) dot plot of AO-stained cells. Cells inside the red area were considered as positive. d Percentage of AO-positive cells. U87 glioma cells treated with Rapamycin 100 nM for 24 h were used as a positive control. Data represent the percentage of AO-positive cells ± SEM;*p < 0.05; **p < 0.01 and ***p < 0.001, using ANOVA (Tukey post-hoc). e Nuclear morphometry of LC3-positive cells. Nuclear Morphometric Analysis (NMA) was performed in LC3-positive cells. The percentage of nuclei in each population of NMA is shown. f Acridine orange intensity ratio calculated to cell populations from Fig. S1A. We compared the intensity of AO staining in cells from the population 1 (cells with normal phenotype based on FSC x SSC) with the intensity of AO staining cells from the population 2 (viable cells with high intracellular complexity) and population 3 (early apoptosis morphology). *p < 0.05, **p < 0.001 - black bar in relation to the correspondent white bar