Human/Mouse NeuroD1 Antibody

NeuroD1 in betaTC-6 Mouse Cell Line.

Neurogenic Differentiation factor 1 (NeuroD1) was detected in immersion fixed betaTC-6 mouse beta cell insulinoma cell line using Human/Mouse NeuroD1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2746) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of NeuroD1 by Immunohistochemistry

Organoid characterization(A) Representative bright-field image of an organoid on the seventh day of culture.(B and C) Immunostaining of a representative organoid for iOSNs (Gap43), HBCs (KRT5), and GBCs (SOX2+, KRT5-). (C) shows high-magnification images of (B).(D–F) Representative immunostaining of organoids for other neurogenic markers.(G) UMAP representation of single-cell RNA-seq of the organoid culture.(H) Stacked violin plots of key marker genes allowing cluster annotation in log scale.(I) Focused UMAP expression plots in the dotted area shown in (G), plotted in log-scale, representing the neurogenic cascade found in the organoids.(J) Histogram plot of the number of distinct ORs found in single cells identified as OSNs based on UCHL1, GAP43, and TUBB3. Image collected and cropped by CiteAb from the following open publication (https://linkinghub.elsevier.com/retrieve/pii/S2667237525000918), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of NeuroD1 by Immunohistochemistry

MCT8-deficient COs present an altered transcriptome.(A) Schematic of the protocol for COs’ maturation using platelet-derived growth factor AA and insulin-like growth factor 1. (B–E) Whole-mount immunolabeling of D65 WT COs containing differentiated NEUROD1+ neurons (B), GFAP+ neural precursor cells and GFAP-AQP4+ astrocytes (C and D), and OLIG2+ oligodendrocyte precursor cells and O4+ premyelinating oligodendrocytes (E); in blue is the nuclear staining with DAPI. B: scale bar 75 μm; C: scale bar 75 μm; inset in C: scale bar 25 μm; D: scale bar 40 μm; E: scale bar 75 μm; inset in E: scale bar 40 μm. Arrows in D indicate AQP4 staining in the membrane of GFAP+ cells. The arrow in the inset in E points to the cellular body of a representative oligodendrocyte; arrowheads point to its cellular processes. (F and G) Volcano plots showing the distribution of DEGs in WT vs. Mut1 (F) and in WT vs. Mut2 (G); each point represents the average of 3 (WT) or 2 (MCT8-deficient) samples consisting of 4 pooled COs for each transcript. (H) Venn comparison of WT vs. Mut1, WT vs. Mut2, and Mut1 vs. Mut2 DEGs. A common cluster (cluster E) between WT vs. Mut1 and WT vs. Mut2 containing 949 DEGs was identified. (I) Venn comparison of the 949 DEGs in cluster E shows similar transcriptomic changes in both MCT8-deficient COs. (J) Heatmap depicting the top 63 DEGs from cluster E. (K) KEGG pathway analysis of the top 63 DEGs in J. (L–O) Heatmaps of T3-regulated genes involved in cerebral cortex development (L), neural cell migration (M), astrocytes and myelination (N), and neurotransmitter receptors, transcription factors, potassium channels, and extracellular matrix protein (O). DEG thresholds (FDR ≤ 0.25 and fold-change ± 4) were identified by gene set–specific analysis in the Partek Flow platform. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/38376950), licensed under a CC-BY license. Not internally tested by R&D Systems.