Fig. 5

AKR1C1-3 levels affect response to vincristine in T-ALL cells. a, b After 48 h from electroporation with two different siRNAs against AKR1C1-3, CCRF-CEM cell lysates were analysed by immunoblotting with AKR1C1, AKR1C2, AKR1C3, and β-Actin-specific antibodies, showing the effective gene silencing of AKR1C enzymes a. Response of AKR1C-silenced CCRF-CEM cells to scalar doses of VCR (48 h) is reported b. c, d AKR1C1-3 activity was stimulated in CCRF-CEM by t-BHQ treatment (5 μM) for 18 h and then evaluated by WB c and calculation of the slope of linear coumberone conversion reaction d. e Dose–response curves of t-BHQ-stimulated and control CCRF-CEM exposed to scalar doses of VCR for 48 h. f Box plot showing the ex vivo response to VCR treatment in terms of cell viability of cells obtained from PPR (n = 4) and PGR (N = 3) T-ALL patient-derived xenografts (PDX-T-ALLs). g Correlation of AKR1C1-3 mRNA expression with the viability of PDTALL cells after ex vivo treatment with 100 nM VCR for 48 h is reported. 95% confidence interval is indicated by dotted lines. All data are expressed as mean ± S.E.M. of three independent experiments. In correlation graphs, Pearson r and relative p values are reported independently for PGR and PPR patient-derived PDX-T-ALLs