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. 2022 Feb 11;149(3):dev200250. doi: 10.1242/dev.200250

Fig. 4.

Fig. 4.

Tcf12-NeuroD1 complex induces active chromatin and expression of neuronal migration genes. (A) Co-IP experiment showing GFP-tagged NeuroD1 interacting with HA-tagged Tcf12 in vitro. This specific interaction was absent in control immunoprecipitates where we co-expressed HA-tagged Tcf12 with only GFP protein. (B) IP experiment showing the specific enrichment of endogenous Tcf12 by NeuroD1 only in the negative control siRNA (left panel). It was absent in Tcf12 knockdown conditions (right panel) during in vitro neurogenesis. (C) IP experiment from the E14.5 mouse cortex showing specific interaction of Tcf12 with NeuroD1 during cortical development in vivo. (D) ChIP qPCR results showing the enrichment of Tcf12 at NeuroD1 target sites after 12 h and 48 h of NeuroD1 induction. (E) ChIP Re-ChIP qPCR experiment showing the NeuroD1 and Tcf12 co-binding at the same target genes. (F) qPCR results showing a change in H3K27ac levels at NeuroD1 target sites upon Tcf12 knockdown. (G) qPCR results showing the expression of NeuroD1 target genes upon Tcf12 knockdown. All the ChIP experiments were performed as three independent biological replicates. The ChIP-Re-ChIP experiment was repeated twice. Data are mean±s.d. Statistical significance was calculated using a paired two-tailed Student's t-test. *P<0.05, **P<0.01.