Fig. 1

Drug response, immunophenotyping and ICD detection in MLH1^−/− cell lines A7450 T1 M1, 328, and 1351. a Dose response curves of MLH1^−/− 328 und MLH1^−/− 1351 cells showing dose-dependent reduction of cell viability. All cell lines were resistant towards CPX (data not shown). Data from MLH1^−/− A 7450 T1 M1 cells are taken from [18]. b Flow cytometric gating strategy and quantitative phenotyping. Cells were treated with chemotherapeutic drugs for 24 h treatment, followed by cell harvest and staining as stated in material and methods. Amounts of cell-surface bound and intracellular molecules were analyzed by multi-color flow cytometry. Results are given as percentage number of cells (%) within 20,000 cells ± standard deviation of three independent experiments each performed in triplicates. c Representative confocal laser scanning microscopy images of MLH1^−/− 328 cells showing increased CalR exposure on the tumor cells’ surface. d Flow cytometric quantification of surface-bound CalR (left graph) as well as quantification of HMGB1 (right graph) in supernatants of MLH1^−/− tumor cells after treatment with different chemotherapeutic drugs. Control cells were left untreated. Experiments were repeated three times each of them performed in duplicates. Values of are given as mean ± SD. *p < 0.05 vs. control; # p < 0.05 vs. 5-FU; § p < 0.05 vs. CPX; one-way ANOVA (Holm Sidak method)