Fig. 5: miR-1298-5p targeted SETD7 in glioma and MSH2 in MDSCs.

a Construction of wild type (WT) and mutant type (MUT) luciferase reporter vectors based on the predicted binding site of miR-1298-5p in SETD7. b 293 T cells were co-transfected with the reporter vectors and miR-1298-5p or miR-Nc. Luciferase activity was assessed 48 h after transfection. c, d The proliferation capacity of U87MG and U251 cells after SETD7 knockdown were assessed using CCK8 assay. e The proliferation capacity of U87MG and U251 cells after SETD7 knockdown were assessed using the Edu assay. f, g Cell cycle analysis for U87MG and U251 cells knocking down SETD7. The percentage of cells arrested in the G1/S phase is analyzed in a histogram. h–k The protein level of SETD7, cyclinD1, P27, p-Akt, Akt in U87MG, U251, and P3 cells knocking down SETD7 were assessed by western blotting. Amounts of protein determined by densitometry of protein bands from three experiments. β-actin was the loading control. l Construction of wild type (WT) and mutant type (MUT) luciferase reporter vectors based on the predicted binding site of miR-1298-5p in MSH2. m 293 T cells were co-transfected with the reporter vectors and miR-1298-5p or miR-Nc. Luciferase activity was assessed 48 h after transfection. n Flow cytometry assay showed that MSH2 knockdown upregulated the proportion of CD14 + HLA-DR low/− MDSCs population. o, p NO and TGF-β in the supernatants of MDSCs were measured after MSH2 knockdown. q, r CD8 + T cell proliferation was determined by flow cytometry 3 days later with CFSE dilution. s, t The protein level of p65 and p-p65 in MDSCs transfected with MSH2 siRNAs were accessed by western blotting. Amounts of protein determined by densitometry of protein bands from three experiments. β-actin was the loading control. Data are shown as the mean ± SD of three independent experiments. Statistical significance was determined using one-way ANOVA test (*P < 0.05; **P < 0.01; ***P < 0.001).