Research Paper Advance Articles

Figure 3. DNMT1 inhibition induces senescence via SPINT2 upregulation. (A–C) HDFs (DT2) were treated with indicated concentrations of 5-AzC for 3 days. (A) Western blot analysis of DNMT1, SPINT2, and p21 protein expression. (B) qRT-PCR analysis of SPINT2 mRNA expression. ^**p < 0.01 vs. DMSO control (V) by Student's t-test. (C) Quantification of SA-β-gal activity ^**p < 0.01 vs. V by Student’s t-test. (D–H) HDFs (DT2) were transfected with siRNAs targeting the indicated genes for 4 days. (D) qRT-PCR analysis of DNMT1 and SPINT2 mRNA levels. ^**p < 0.01 vs. negative control siRNA (NC) by Student’s t-test. (E) Western blot analysis of DNMT1, SPINT2, and p21 protein expression. (F) Quantification of SA-β-gal activity. ^**p < 0.01 vs. NC by Student’s t-test. (G) Western blot analysis of DNMT1, SPINT2, and p21 protein expression. (H) Quantification of SA-β-gal activity following co-transfection ^**p < 0.01 vs. NC by Student’s t-test. (I, J) HDFs (DT2) were infected with the recombinant lentivirus encoding SPINT2 cDNA for 4 days. GFP-encoded recombinant lentivirus was used as a control. (I) Western blots analysis. (J) Quantification of SA-β-gal activity. ^**p < 0.01 vs. GFP by Student’s t-test.