Research Paper Volume 11, Issue 24 pp 12581—12599

FOXP3 pathogenic variants cause male infertility through affecting the proliferation and apoptosis of human spermatogonial stem cells

Figure 5. Influence of FOXP3 Knockdown on the proliferation, DNA synthesis and apoptosis of human SSC line. (A) Fluorescence microscope and phase-contrast microscope revealed transfection efficiency of FOXP3-siRNAs using the FAM-labeled miRNA oligonucleotides. Scale bars = 40 μm. (B) Western blots showed the protein changes of FOXP3 by FOXP3-siRNAs in human SSC line. (C) Real-time PCR displayed the mRNA changes of FOXP3 by FOXP3-siRNAs in human SSC line. (D) EDU incorporation assay showed the percentages of EDU-positive cells affected by control siRNA, FOXP3-siRNA 1 and FOXP3-siRNA 3 in human SSC line. Scale bars = 40 μm. Values are means ± SD from three independent experiments. (E) CCK-8 assay demonstrated the proliferation of human SSC line after transfection with FOXP3-siRNA 1 and FOXP3-siRNA 3. (FG) Annexin-V/PI staining and flow cytometry displayed the percentages of early apoptosis and late apoptosis of human SSC line transfected with FOXP3-siRNA 1 and control siRNA. All values are means ± SD from three independent experiments. * indicated statistical significance (p<0.05) between FOXP3-siRNAs and control siRNA treatments. All experiments were repeated for at least three times.