Fig. 1
Effect of INO, GO and γ-calicheamicin on cell viability, cell-cycle perturbation and G2/M checkpoint activation in ALL and AML cell lines. A Reduction of the cell viability of ALL (blue) and AML (red) cell lines treated for 24, 48 and 72 h with increasing concentration of INO and GO, respectively. Each point represents the mean ± standard deviation of normalized cell viability obtained from at least three independent experiments. Dashed lines represent the indicative values of inhibitory concentration 50 (IC50). B Expression of CD33 and CD22 in AML (red) and ALL (blue) cell lines by flow cytometry. Histograms represent the mean and standard deviation of the mean fluorescence intensity (MFI) of AML and ALL cell lines obtained from at least three independent experiments. The table reports the Pearson correlation coefficient in AML and ALL cell lines between CD22 expression and IC50 values after 24, 48 and 72 h of GO and INO, respectively. C Reduction of the cell viability of ALL (blue) and AML (red) cell lines treated for 24, 48 and 72 h with increasing concentrations of γ-calicheamicin. Each point represents the mean ± standard deviation of normalized cell viability obtained from at least three independent experiments. Dashed lines represent the indicative values of inhibitory concentration 50 (IC50). D Cell-cycle profile analysis of ALL and AML cell lines treated for 18, 24 and 48 h with subtoxic concentrations of INO (near IC50 values after 24 h) and GO (near IC50 values after 24 h), respectively. E, F Representative immunoblotting analysis of ALL and AML cell lines treated with INO (near IC50 values after 24 h) or GO (near IC50 values after 24 h) for 18 h. β-actin was used for loading normalization. G Histograms representing the mean ± standard deviation the relative bands intensity of CCNB1, phospho-CDK1Tyr15 and phospho-H2AXSer139 proteins in ALL (blue) and AML (red) cell lines treated INO or GO for 18 h. H Immunoblotting analysis of ALL and AML cell lines treated with γ-calicheamicin (IC50 values after 24 h) for 18 h. β-actin was used for loading normalization. I Histograms represent the mean and standard deviation the relative bands intensity of phospho-ATMSer1981, phospho-ATRSer428, phospho-CHK2Thr68, phospho-CHK1Ser345 and phospho-WEE1Ser642 proteins in ALL (blue) and AML (red) cell lines treated INO or GO for 18 h. All error bars represent the mean ± SD