Fig. 3.

pCYB5Aalt Nalm 6 showed a distinct transcriptional program and resistance to Venetoclax induced cell death. A CYB5Aalt-ORF is stably overexpressed in NALM-6 cells. NALM-6 cells were transfected with pCYB5Aalt (+) or pEmpty (−). RT-PCR was used to confirm overexpression of CYB5Aalt-ORF mRNA. Two representative samples of each are shown. GAPDH was used as a control. NTC = non-template control. Full length electrophoretic gel is displayed in Additional file 17. B pCYB5Aalt NALM-6 show a distinct transcriptional program compared to empty vector controls. Samples were grouped according to the 500 most variably expressed genes between samples. Columns indicate pCYB5Aalt clones and empty vector controls, rows represent gene expression in rlog for each sample. Colour of the heatmap indicates relative expression strength according to the deviation from the mean of all samples in rlog. C Viability of pCYB5Aalt NALM-6 cells and pEmpty upon Venetoclax treatment (n = 3). Cells were incubated with different concentrations of Venetoclax (0 μM, 0.2 μM, 0.5 μM, 5 μM, 10 μM) for 72 h. Viability was assessed photometrically after 3 h incubation time with WST-1. Absorbance values were normalised to untreated control samples and shown as viability (%). Bars represent mean viability, error bars ±standard deviation of three independent experiments each with five technical replicates per concentration. Mann-Whitney-U-Test, ***P-value< 0.001, ns = non-significant. D Cell death of pCYB5Aalt NALM-6 and pEmpty upon Venetoclax treatment (n = 4). Cells were incubated at different concentrations of Venetoclax (0 μM, 0.2 μM, 0.5 μM, 5 μM, 10 μM, 20 μM) for 48 h. Cell death was assessed by staining cells with PI. Bars represent percentage of PI positive cells, error bars ±standard deviation of four independent experiments each with technical duplicates for each concentration. Mann-Whitney-U-Test, ***P-value< 0.001, **P-value< 0.01, *P-value< 0.05, ns = non-significant