Journal: Development (Cambridge, England)

Article Title: Tcf12 and NeuroD1 cooperatively drive neuronal migration during cortical development

doi: 10.1242/dev.200250

Figure Lengend Snippet: Dynamics of the H3K27ac mark following NeuroD1 expression. (A) Schematic of analysing H3K27ac dynamics upon NeuroD1 expression. (B) Global dynamics of H3K27ac enrichment before and after NeuroD1 induction (12 h and 48 h). R1 and R2 are replicates. (C) Boxplots show average H3K27ac enrichment in each cluster (C1 to C7). Middle bars show median values, boxes show first to third interquartile ranges, whiskers show the minimum and maximum of data range and beyond that are outliers. (D) NeuroD1-bound and transcriptionally induced sites. (E) Motif enrichment profiles at transcriptionally active and inactive sites.

Article Snippet: The eluted fractions were immunoblotted with Tcf12 antibody (SCBT sc-28364 X) and NeuroD1 antibody (CST 4373S) (1:1000).

Techniques: Expressing

Journal: Development (Cambridge, England)

Article Title: Tcf12 and NeuroD1 cooperatively drive neuronal migration during cortical development

doi: 10.1242/dev.200250

Figure Lengend Snippet: Identification of potential NeuroD1 co-factors. (A) Schematic of ISMARA analysis for co-enriched TF selection. (B) Overlap of upregulated motifs predicted from ISMARA with bHLH transcription factors. (C) Activity profiles for selected motifs from the overlap of ISMARA motifs and bHLH factors. R1 and R2 are the replicates for each time point (0 h, 12 h and 48 h). Data are mean±s.d. (D) Expression of selected factors in different cortical layers. (E) In situ hybridisation images (Allen Brain Atlas) showing the expression of NeuroD1 and Tcf12 in developing cortex at E11.5, E13.5 and E15.5. (F) Bar chart showing the expression level of NeuroD1 in VZ, SVZ and CP regions. (G) Bar chart showing the expression level of Tcf12 in VZ, SVZ and CP regions. (H) Bar chart showing the expression level of Tcf12 in proliferating progenitors (PP), differentiating progenitors (DPs) and neurons (NNs). Data are mean±s.d. Statistical significance was calculated using a paired two-tailed Student's t -test. * P <0.05, ** P <0.01, *** P <0.001.

Article Snippet: The eluted fractions were immunoblotted with Tcf12 antibody (SCBT sc-28364 X) and NeuroD1 antibody (CST 4373S) (1:1000).

Techniques: Selection, Activity Assay, Expressing, In Situ, Hybridization, Two Tailed Test

Journal: Development (Cambridge, England)

Article Title: Tcf12 and NeuroD1 cooperatively drive neuronal migration during cortical development

doi: 10.1242/dev.200250

Figure Lengend Snippet: NeuroD1 and Tcf12 are co-expressed in distinct subpopulations during cortical development. (A) Expression of NeuroD1 and Tcf12 in different brain cell types in E14.5 cortex shows the highest NeuroD1 expression in the migrating neuronal population and the Tcf12 expression levels. (B) Pseudo-timecourse trajectory of SVZ migrating cells. Numbers show the clusters arranged in developmental pseudotime. (C) Expression of progenitor, neurogenic and neuronal markers in a SVZ migrating subpopulation arranged by pseudotime, where we see increasing NeuroD1 expression levels.

Article Snippet: The eluted fractions were immunoblotted with Tcf12 antibody (SCBT sc-28364 X) and NeuroD1 antibody (CST 4373S) (1:1000).

Techniques: Expressing

Journal: Development (Cambridge, England)

Article Title: Tcf12 and NeuroD1 cooperatively drive neuronal migration during cortical development

doi: 10.1242/dev.200250

Figure Lengend Snippet: 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.

Article Snippet: The eluted fractions were immunoblotted with Tcf12 antibody (SCBT sc-28364 X) and NeuroD1 antibody (CST 4373S) (1:1000).

Techniques: Expressing, Migration, Co-Immunoprecipitation Assay, In Vitro, Control, Negative Control, Knockdown, In Vivo, ChIP-qPCR, Binding Assay, Two Tailed Test

Journal: Development (Cambridge, England)

Article Title: Tcf12 and NeuroD1 cooperatively drive neuronal migration during cortical development

doi: 10.1242/dev.200250

Figure Lengend Snippet: A schematic showing how the functional cooperativity of Tcf12 and NeuroD1 in specific subpopulations of the developing cortex creates the gene regulatory program essential for neuronal migration. Both Tcf12 and NeuroD1 are highly co-expressed during the transition of cortical progenitors from proliferative to neurogenic divisions. During this phase, Tcf12 forms a complex with NeuroD1 and co-occupies a subset of NeuroD1 target loci. This Tcf12-NeuroD1 cooperativity is essential for a gain in active chromatin and expression of neuronal migration genes, and therefore for the correct migration of newborn neurons.

Article Snippet: The eluted fractions were immunoblotted with Tcf12 antibody (SCBT sc-28364 X) and NeuroD1 antibody (CST 4373S) (1:1000).

Techniques: Functional Assay, Migration, Expressing

Journal: BMC Neuroscience

Article Title: Sonic hedgehog (Shh)/Gli modulates the spatial organization of neuroepithelial cell proliferation in the developing chick optic tectum

doi: 10.1186/1471-2202-13-117

Figure Lengend Snippet: Radial organization and immunocytochemical patterns at the end of DS1. A . Ectopic expression of pShhiEGFP in an electroporated OT (ED4.5); patches of positive NE cell bodies at the ventricular zone (VZ) and of postmitotic neurons at the premigratory zone (PMZ) can be observed. B . Hematoxylin-Eosin staining. Arrowheads point to mNE cells located along ventricular zone; they show PH3 nuclear labeling ( C ) and Notch reactive cytoplasm ( D ). Arrows point to neurons in the premigratory zone; they display nuclear NeuroD reactivity ( E ) and βIIITub reactive perikarya ( F ). Bar: 10 μm.

Article Snippet: Rabbit polyclonal anti-neurod (NeuroD1) , Lifespan Biosciences , , , Synthetic peptide DDDQKPKRRGPKKKKM Conjugated to malemide-activated KLH Human NeuroD1 (aa 76–91) , 1:100.

Techniques: Expressing, Staining, Labeling

Journal: BMC Neuroscience

Article Title: Sonic hedgehog (Shh)/Gli modulates the spatial organization of neuroepithelial cell proliferation in the developing chick optic tectum

doi: 10.1186/1471-2202-13-117

Figure Lengend Snippet: Primary antibodies characteristics

Article Snippet: Rabbit polyclonal anti-neurod (NeuroD1) , Lifespan Biosciences , , , Synthetic peptide DDDQKPKRRGPKKKKM Conjugated to malemide-activated KLH Human NeuroD1 (aa 76–91) , 1:100.

Techniques: Translocation Assay, Recombinant