Eif2s3y Promotes the Proliferation of Spermatogonial Stem Cells by Activating ERK Signaling

Figure 4

Depletion of Eif2s3y resulted in proliferation abnormality in goat SSCs. (a, b) The schematic of lentivirus plasmid CD513B-U6-MCS and CD513B-U6-shEif2s3y. (c) Typical images of shControl (left) and shEif2s3y (right) SSCs. Scale bar, 200 μm. (d) Proliferation curve of shControl and shEif2s3y SSCs and the results of population doubling time (PDT) determination. (e) EdU incorporation assay of shControl (up) and shEif2s3y (down) SSCs. Scale bar, 400 μm. (f) The ratio of EdU-positive cells to total cells. Data are presented as and are represented by three independent repetitions. (g) RT-PCR analysis of the expression levels of Eif2s3y, Pcna, Cyclin D, and Zbtb16 in shControl and shEif2s3y SSCs in vitro. (h) Western blotting detected the protein expression of ZBTB16, eIF2γ, PCNA, and Cyclin D in shControl and shEif2s3y SSCs. GAPDH was used as a loading control. (i) Gray intensity analysis of WB results normalized to GAPDH in (h). Data are presented as and are represented by three independent repetitions; , , and .

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