Fig. 5.

SALL3 interacts with DNMT3B and modulates DNMT function. a Lysate prepared from 253G1 cells was immunoprecipitated with anti-SALL3 or normal rabbit IgG. The immunoblot was analyzed with anti-DNMT3A, DNMT3B, and DNMT1 antibodies. Molecular weight is indicated as Mr (k). b Lysate prepared from 253G1 cells was immunoprecipitated with anti-DNMT3B or normal sheep IgG. The immunoblot was analyzed with anti-SALL3 antibody. c In vitro DNMT activity assay. DNMT activity of the nuclear fractions prepared from 253G1 (WT) cells and the 253G1 SALL3^−/− cells was measured (n = 3, biological replicates). ^*P < 0.01, two-sided t test. d qRT-PCR analysis of neural cell markers PAX6 and SOX1. Total RNA was isolated from 253G1-derived (WT), 253G1 SALL3^−/−-derived and 253G1 SALL3^−/− DNMT3B^−/mut-derived neural cells (n = 6, biological replicates). ^*P < 0.01, one-way ANOVA with post hoc Tukey–Kramer test. e qRT-PCR analysis of cardiomyocyte markers GATA4, NKX2.5, and TNNT2. Total RNA was isolated from 253G1-derived (WT), 253G1 SALL3^−/−-derived and 253G1 SALL3^−/− DNMT3B^-/mut-derived cardiomyocytes (n = 6, biological replicates). ^*P < 0.01, one-way ANOVA with post hoc Tukey–Kramer test. Error bars represent mean ± SD