Fig. 3
From: RETRACTED ARTICLE: REST regulates the cell cycle for cardiac development and regeneration
REST binds p21 to repress its expression. a, b Immunostaining (n = 4/group) showing upregulated p21 protein level in the Rest deleted E12.5 and P3 hearts. Scale bar = 40 µm. c Top, Sequence alignment of p21 between mouse and human indicating a conserved RE-1 motif (red vertical line) located at the 3′ UTR. Bottom, The p21-RE1 motif and its mutated (Mut) or deleted (Del) counterpart. d Luciferase reporter assays with a 690-bp mouse DNA fragment containing the wild type (WT), mutated, or deleted RE1 motif reveals that the intact p21-RE1 motif is required for repressing the reporter gene transcription in primary cultured P1 mouse cardiomyocytes. e qChIP using REST antibodies indicates enrichment of p21-RE1 motif containing DNA fragments in the ventricles of E12.5 and P1, but not P21. f qChIP with P2 ventricles using the REST and histone mark antibodies showing the REST-dependent histone modification of the p21-RE1 motif. n = 3/group for d–f, mean ± s.d., *p < 0.05 by one-way ANOVA followed by Tukey’s test in d; unpaired Student’s t test in b, e, f. NS no significance
