Figure 3.

Chromomycins A_5-8 induce autophagy. B16-F10 melanoma cells were treated with DMSO (C-), as a negative control, doxorubicin (Dox), as a positive control, and CA_5-8 (CA5-8) for 24 h. (A) Representative contour plot graphs of cells with granularity gated on high side scatter (SSC) and average forward scatter (FSC) region by flow cytometry and (B) Expression of LC3BII and LC3BI obtained by Western blot and (C), column graph of Western blot analysis. (D), Representative dot plots of acridine orange (AO) staining. Acidic vesicular organelles (AVOs) were gated on the region with increased red fluorescence by flow cytometry and (E) the graph showing the percentage of AVOs. Data presented in graphs as mean ± standard deviation of 3 independent experiments. The AVOs detection was performed in triplicate. The p values of C- compared to the treated groups are above each treated group. **p=0.01 and ***p=0.001.