Fig. 3

Primary CD3, CD4 and CD8 T lymphocytes purified from HDs’ peripheral blood were transduced by the LV-miR-24-3p construct containing GFP. A miR-24-3p expression levels after transduction were determined by qRT‒PCR in RNA extracted from the lenti-miR-ctrl (black bar) and lenti-miR-24-3p groups (n = 5). CD3 T cells were transduced with an efficiency of 93–94%, as shown by flow cytometry. Lentiviral transduction increased miR-24-3p expression in CD3 T cells. B Apoptosis was measured by FACS (cells were labeled with annexin-V-FITC and PI). The overexpression of miR-24-3p induced apoptosis in T cells (CD3, CD4 and CD8) compared to the negative control [CD3 (*** n = 5, p < 0.0002), CD4 (n = 5 ***p < 0;0001), CD8 (n = 5 ** p < 0.002)]. C Primary CD3 T lymphocytes purified from peripheral blood from AML patients were transduced with the LV-anti-miR-24-3p construct containing GFP, and the expression of GFP after transduction was measured by FACS (94%) (mean ± SEM, n = 5). The inhibition of miR-24-3p expression by anti-miR-24-3p decreased cell apoptosis (cells were labeled with annexin-V-FITC and PI) compared to that of the scrambled anti-miR control group. All values are shown as the mean ± SEM, n = 5. *p < 0.05; **p < 0.001, *p < 0.02. D RT‒qPCR analysis of the immunosuppressive, antiapoptotic and proapoptotic genes. E Western blot analysis of the antiapoptotic proteins Bcl-2 and Xiap and the proapoptotic proteins caspase-9 and caspase-3 after transduction of CD3 T cells from HDs by the LV-miR-24-3p construct containing GFP or the lenti-CTRL-GFP construct and CD3 T cells from AML patients