Fig. 1

Induction of apoptosis and phenotypic alterations in NSCLC cells by classic chemotherapeutics. a Effect of chemotherapeutics in the number of A549 cells after 48 h of treatment. C – control (DMSO not exceeding 0.1%); Cis – Cisplatin 40 μM; Eto – Etoposide 13.2 μM; Carb – Carboplatin 200 μM; Pac – Paclitaxel 100 nM; Gem – Gemcitabine 0.96 μM; Cis + Eto – Cisplatina 40 μM + Etoposide 13.2 μM; Carb+Pac – Carboplatin 200 μM + Paclitaxel 100 nM. Cell number was analyzed by cell counting in a flow cytometer. Data represent the mean ± SEM;*p < 0.05 and **p < 0.01 in relation to the control considered as 100%, using ANOVA (Tukey post-hoc). b Representative images of cells treated with chemotherapeutics for 48 h. Images were obtained by phase-contrast inverted microscope (200x). c Effect of chemotherapeutics on the induction of apoptosis and necrosis after 48 h in A549 cells. Dot blot graphs show annexin V-FITC/ PI co-staining with quadrant analysis, as following: early apoptotic cells (upper left), late apoptotic cells (upper right), plasma membrane permeabilization (low right). Results are expressed as the percentage of cells in each quadrant ± SEM; *p < 0.05 and **p < 0.01 in relation to the control, using ANOVA (Tukey post-hoc). C+: A549 cells treated with Cis 80 μM. d Levels of active caspase 3 measured through flow cytometry. Data represent the mean ± SEM;*p < 0.05 and **p < 0.01 in relation to the control considered as 100%, using ANOVA (Tukey post-hoc). Dot plots are shown in Fig. S1D. C+: A549 cells treated with Cis 80 μM. e Nuclear Morphometric Analysis. Dot blots represent Nuclear area versus Nuclear Irregularity Index (NII). Dots in each condition correspond to single nuclei (at least 50 nuclei are shown to each condition). Nuclear phenotype populations are shown in the legend on the bottom, and the percentage of nuclei in each population is shown in the pie chart. Representative figures of DAPI-stained nuclei are shown in Fig. S1C