Figure 5 | Scientific Reports

Figure 5

From: Single cell RNA-sequencing identified Dec2 as a suppressive factor for spermatogonial differentiation by inhibiting Sohlh1 expression

Figure 5The alternative text for this image may have been generated using AI.

DEC2 transcriptionally suppresses Sohlh1 and subsequently silences Kit expression both in vitro and in vivo. (a) Effects of Dec2 knockdown (shDec2) and add-back (shDec2 + F-DEC2r) on the expression levels of selected marker genes in SSC culture. Five days after shRNA-containing lentivirus infection, cultured SSCs were harvested and subjected to RT-qPCR. (b) Effect of simultaneous knockdown of Dec2 (shDec2) and Sohlh1 (shSohlh1) on the expressions of Dec2, Sohlh1, and Kit mRNAs in SSC culture. In (a,b), the vertical axes indicated the relative mRNA expression levels (CTR = 1). Dunnett’s test was performed against shDec2 samples. (c) Effects of the combination of shDec2, F-DEC2r, and shSohlh1 on the number of KIT(+) cells in SSCs. Dunnett’s test against shDec2 samples was performed for statistical analysis. (d) Effect of shDec2 on KIT expression level in P3.5 germ cells. KIT intensity was measured by flow cytometry at three days after shDec2 lentivirus infection. The vertical axes indicated the relative KIT intensity (CTR = 1). Cell sorting scheme and raw flow cytometry data are shown in Fig. S12. Student’s t-test was performed for statistical analysis. All the experiments were performed by three biological replicates. Error bars indicated standard deviations. *p value < 0.05; **p value < 0.01; ***p value < 0.001.

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