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. 2023 Nov 17;24(22):16472. doi: 10.3390/ijms242216472

Figure 2.

Figure 2

ShRNA-dependent knockdown of MEN1 decreases fitness and induces apoptosis in B-ALL cell lines independently of the presence of MLLr. (A–C) B-ALL cell lines were transduced with shRNAs targeting MEN1 (shRNA#5 or shRNA#6) or control vectors (scr) expressing the fluorescent marker RFP. (A) On day 4, RFP+ cells were sorted and MEN1 expression was measured by immunoblot. TUBB was used as loading control. (B) MEN1 knockdown decreases fitness of B-ALL cell lines. Starting from day 4 after transduction (day 0) the percentage of the RFP+ cells was measured by flow cytometry. The data are shown as percentage of RFP+ cells (mean ± SD N = 3). * = p < 0.05, ** = p < 0.01, *** = p < 0.005 (C) MEN1 knockdown induces apoptosis in B-ALL cell lines. RFP+ cells were sorted on day 3 after transduction and incubated under the normal conditions for 72 h. Apoptosis was measured by Annexin V/7-AAD staining. The data are shown as specific apoptosis (SA). SA was calculated as: SA%, =100 × (AE − AC)/(100 − AC), where AE and AC equals the percentage of apoptotic cells in the experimental and control groups, respectively (mean ± SD N = 3). (D) NALM6 and SUP-B15 cell lines are not sensitive to VTP50469. The sensitivity was measured by 6 days MTT metabolic test. The data are shown as percentage to control treated with vehicle DMSO (mean ± SD, N = 3).