Fig 2: Knockdown of SETDB1 altered cell adhesion and cell spread. (A) Morphology of siSETDB1 and siCtrl cells. Scale bar = 100 µm. (B) Detection of cell spreading area by F-actin (red) and DAPI (blue) staining. Scale bar = 100 µm. (C) Analysis of cell area is shown in the column diagram. (D) Morphology of cells adhered for 30 min. Scale bar = 100 µm. (E) Detection of adherent cells in each group by CCK-8 assay. Significance was presented as * p < 0.05, *** p < 0.001. Error bars represented the standard error of the mean (SEM).

Fig 3: SETDB1 expression in porcine testes. (A–C) Immunofluorescence panel of SETDB1 (red), H3K9me3 (green), UCH-L1 (green), SYCP3 (green), and DAPI (white) at 7, 60 and 150 days. Scale bar = 100 µm. (D) The proportion of SETDB1+ cells in undifferentiated spermatogonia (UCH-L1+). (E) The proportion of SETDB1+ cells in spermatocytes (SYCP3+). Significance was presented as * p < 0.05 or *** p < 0.001. Error bars represented the standard error of the mean (SEM).

Fig 4: Knockdown of SETDB1 in porcine SSCs. (A) Immunofluorescence panel of VASA (red), PLZF (green), UCH-L1 (red), SV40 (green), PCNA (red), negative control (IgG; green) and DAPI (blue) in porcine SSCs. Scale bar = 50 µm. (B) qRT-PCR analysis of the relative mRNA level of SETDB1 in porcine SSCs. (C) Western blot analysis of the protein levels of SETDB1 and ACTB in porcine SSCs. (D) The protein level of SETDB1 relative to ACTB. Significance was presented as *** p < 0.001. Error bars represented the standard error of the mean (SEM).

Fig 5: SETDB1 regulated MMP3/10 expression in porcine SSCs. (A) qRT-PCR analysis of the relative mRNA level of MMP3/10 in porcine SSCs. (B) Western blot analysis of the protein levels of MMP3/10 and ACTB in porcine SSCs. The protein level of MMP3/10 relative to ACTB is shown in the column diagram. (C) Western blot analysis of the protein levels of H3K9me3 and histone H3 in porcine SSCs. The protein level of H3K9me3 relative to histone H3 is shown in the column diagram. Significance was presented as * p < 0.05, ** p < 0.01 or *** p < 0.001. Error bars represented the standard error of the mean (SEM).