huc elavl3 antibody  (Proteintech)

Journal: bioRxiv

Article Title: The RNA and protein landscapes of mouse brain organoids

doi: 10.64898/2026.03.13.711293

Figure Lengend Snippet: (A) Schematic timeline of brain organoids generation from mESCs. mESCs and brain organoids developed for 7, 14 and 21 days were profiled by transcriptomics and proteomics. Part of the figure was created with Biorender. (B) Immunostaining of mESCs-derived brain organoids. Cultures of mESCs were stained with NANOG (green), POU5F1/OCT4 (red), and nuclei with DAPI (blue). Scale bars: 10 µm. (C) Organoid cryosections were stained with NESTIN and PAX6 (at day 7), ELAVL3 and TBR1 (at day 14), and TUBB3 and GFAP (at day 21). Scale bars: 100 µm. (D) PCA showing the distribution of the transcriptomes of mESCs, brain organoids (at days 7, 14, and 21), and NBB samples. (E) Venn diagram showing the overlapping upregulated and downregulated genes between D21 organoids and NBB, both compared to mESCs. (F) Histograms of enriched GO terms in NBB vs ESCs and D21 organoids vs ESCs performed on upregulated genes (black and grey bars) and on downregulated genes (red and light red bars) determined using DAVID. For each term category (Cellular component, molecular function, and biological process), the five highest enriched terms in each gene list are shown.

Article Snippet: The primary antibodies were (species, provider; catalog number): anti-NESTIN –Rat-401- (mouse, Santa-Cruz, sc-33677); PAX6 (Rabbit, Covance, PRB-278P); NANOG (mouse, BD Pharmingen, 560259); POU5F1 (rabbit, Cell Signalling, 2840), ELAVL3 (mouse, Santa Cruz, sc-515624), TUBB3 (mouse, Covance, MMS-435P); TBR1 (rabbit, Cell Signalling, 49661), REELIN (mouse, Covance; MAB5364), GRM5 (rabbit, Millipore, AB5675), and GFAP (rabbit, Dako, Z0334).

Techniques: Transcriptomics, Immunostaining, Derivative Assay, Staining

Journal: bioRxiv

Article Title: Nonsense-mediated decay masks cryptic splicing events caused by TDP-43 loss

doi: 10.1101/2025.07.09.664014

Figure Lengend Snippet: a. Sashimi plots show examples of TDP-43 regulated cryptic splicing events across different conditions. The “TDP-43 sites” track marks the observed TDP-43 binding . Gene structure is shown at the bottom. b. RT-PCR confirms that NMD inhibition by 11j treatment in iNeurons (top panel) or iMNs (bottom panel) with TDP-43 knockdown increased the level of the CELF5 cryptic isoform. c. Sashimi plots show cryptic splicing in ELALV3 across conditions. Details, as in a . d. RT-PCR confirms that NMD inhibition by 11j treatment in the TDP-43 knockdown background increased level of the cryptic isoform of ELALV3 in iNeurons (top panel) and iMNs (bottom panel). e. Western blots show that TDP-43 knockdown reduced full-length ELAVL3 protein levels in iNeurons. f. Sashimi plots show cryptic splicing in DNMT3A across conditions. Details, as in a . g. RT-PCR confirms that only combined NMD inhibition by11j treatment and TDP-43 knockdown unmasked the cryptic isoform of DNMT3A in iNeurons (top panel) and iMNs (bottom panel). h. Western blots show that TDP-43 knockdown reduced full-length DNMT3A protein levels in iNeurons. Unless otherwise stated, quantitation ( n = 3 per condition) is presented as mean values +/- SEM; p values were two-sided Welch’s t-test; ns (not significant), p > 0.05; *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ****, p ≤ 0.0001.

Article Snippet: Membranes were blocked in EveryBlot Blocking Buffer (Bio-Rad, 12010020) for 1 h then incubated overnight at 4°C in blocking buffer containing antibodies against phosphorylated-UPF1 (1:1,000; EMD Millipore, 07-1016 ), UPF1(1:10,000; Abcam, ab109363), TDP-43 (1:1,500; Proteintech, 10782-2-AP), ELAVL3 (1:1,000; Proteintech, 55047-1-AP), DNMT3A(1:500; Abcam, ab2850), Beta Tubulin (1:40,000; Proteintech, 66240-1-Ig), GAPDH (1:2,000; Sigma-Aldrich, G8795), Histone H3 (1:5,000; Abcam, ab1791).

Techniques: Binding Assay, Reverse Transcription Polymerase Chain Reaction, Inhibition, Knockdown, Western Blot, Quantitation Assay