Fig. 7
Cellular and molecular markers of apoptotic cell death on NSCLC cells incubated with CIGB-300 in concentration-dependent and time-course experiments. (a) Representative flow cytometry dot plots, upper panel: X-axis AnnexinV-Fluorescein(FITC) vs. Y-axis (PI) and lower panel: X-axis FSC vs. Y-axis SSC, obtained after AnnexinV-PI staining of A549 cells incubated with CIGB-300 (IC90) during indicated time points. Percent of cell populations is indicated within each quadrant, for Q2 mean ± SD of three replicates; veh, vehicle. (b) Summary of the AnnexinV-PI staining experiments on the four cells lines, at a fixed time point of 24 h using two peptide concentrations (IC50 & IC90). Here, AnnexinV + comprised both A+/PI + and A+/PI- cell populations. (c) WB for detection of PARP cleavage after incubation of A549 or SK-MES-1 cells with CIGB-300 (IC50) during 3–6 h. B-actin protein was used to normalize total protein content/lane; +, CIGB-300; CX, CX-4945; veh, vehicle incubated cells, for CIGB-300 (PBS1X) and for CX-4945 (DMSO). Molecular weights of full length PARP (118 kDa), expected PARP cleavage product (89 kDa) and B-actin (43 kDa). Blots strips in (c) were cropped from different parts of the same membrane as indicated by white in-between lines. Images are representative of two independent experiments. Raw images can be found at Supplementary Information File#2.
