Search

Additional file 6: Fig. S6 Correlation matrix to all measurements assessed. ρ coef. Represent the pearson correlation coefficients. Red and green boxes correspond to all negative or positive significant correlations, respectively. Abbreviatures: ‘% Annex+/PI- cells’ - % of annexin+/Propidium Iodide negative cells; ‘% CRT+ cells’ - % of calreticulin-positive cells; ‘% AO+ cells’ - % of acridine orange-positive cells; ‘ATP’ - levels of extracellular ATP; ‘HMGB1’ - levels of extracellular HMGB1; ‘CRT Marg.’ - Margination of Calreticulin; ‘%LR, %N, %SR and %Irreg’ - percentage of ‘Large and Regular’, ‘Normal’, ‘Small and Regular’ and ‘Irregular’ nuclei from NMA analysis; ‘Nuc Area’ - nuclear area.

Additional file 4: Fig. S4 Calreticulin (CRT) exposure and its correlation with cell area. (A) Representative histograms of cell count and CRT exposure in the cell surface, as obtained by flow cytometry. *p

Additional file 7: Fig. S7 Influence of HMGB1 levels in the prognosis of NSCLC. Kaplan–Meier survival analysis according to HMGB1 levels.

Additional file 3: Fig. S3 Nuclear and LC3 co-staining. Representative images from each treatment are shown. Double arrowheads: nuclei classified as small and regular in the NMA; Arrows – nuclei classified as large in the NMA; Single arrowhead: nuclei classified as normal in NMA.

Additional file 5: Fig. S5 Influence of CRT levels in the prognosis and staging of NSCLC. (A) Kaplan-Meier survival curve to patients expressing high or low levels of CRT in the TCGA cohort. (B) Dead/live ratio of patients for grade i-iv tumors according to CRT levels. (C to F) Influence of CRT levels on the outcome (i.e. dead or alive) of patients with stage i to iv NSCLC.

Additional file 2: Fig. S2 Morphological changes and caspase activation in A549 cells treated with chemotherapeutics. (A) Analysis of cell phenotype assessed by flow cytometry. Population 1 indicates normal cell morphology. Reduced cell size (low FSC) with increased intracellular granularity (high SSC) is suggestive of early apoptosis (population 3); high intracellular granularity (high SSC) suggests cytoplasmic vacuolization, as observed after autophagy activation (population 2); and cell debrie suggests late apoptosis, which is characterized by cell fragmentation (population 4). A scheme showing these populations is shown on the right. (B) Cell area measured through images from immunocytochemistry. (C) Representative images of nuclear staining with DAPI (magnification: 200x). (D) Representative plots of active caspase 3 versus FSC. Red area delimits the population of cells with small size (i.e. cell shrinkage, a typical change of early apoptotic cells) and active caspase 3. Numbers represent the percentage of cells inside the red area ± SEM. Cisplatin 80 μM was used as a positive control (C+).

Additional file 1: Fig. S1 Dose-response curves in A549 cells. Cells were treated with the chemotherapeutics as indicated, for 48 h. After this, cell number was determined through flow cytometry.