MKK7-mediated phosphorylation of JNKs regulates the proliferation and apoptosis of human spermatogonial stem cells

Figure 2 The influence of mitogen-activated protein kinase kinase 7 knockdown on human spermatogonial stem cell proliferation. A: Quantitative real time PCR revealed mitogen-activated protein kinase kinase 7 (MKK7) mRNA level changes in a human spermatogonial stem cell (SSC) line after treatment with MKK7-small interfering RNA (siRNA) 1-, 2- and 3; B and C: Western blotting showed MKK7 protein level alterations in the human SSC line after the transfection of MKK7-siRNA 1-, 2- and 3. Beta actin (ACTB) was used as the loading control for the total protein; D: Cell counting kit (CCK)-8 assay illustrated the proliferation of human SSCs transfected with the Control-siRNA and MKK7-siRNA 3; E and F: The relative levels of proliferating cell nuclear antigen (PCNA) protein in human SSCs after transfection with the Control-siRNA and MKK7-siRNA 3; G and H: The percentages of EdU-positive cells in human SSCs transfected with the Control-siRNA and MKK7-siRNA 3. Scale bars: G, 50 μm. ^aP < 0.05 denotes a significant difference between the MKK7-siRNA 3 and Control-siRNA groups. DAPI: 4,6-diamidino-2-phenylindole.