rabbit anti tubb3  (Cell Signaling Technology Inc)

Journal: Advanced Science

Article Title: TTNPB Promotes Human Pluripotent Stem Cell‐to‐Neural Stem Cell Transition via Modulation of Chromatin Accessibility and the S‐(5′‐adenosyl)‐L‐homocysteine/Choline Metabolic Network

doi: 10.1002/advs.202515648

Figure Lengend Snippet: NSCs and ANSCs transplantation ameliorates depressive‐like behaviors and remodels hippocampal transcriptomes in LPS‐ and CUMS‐induced rat models. (A), Schematic illustration of the experimental design for the establishment of LPS‐ and CUMS‐induced depressive‐like rat models and stereotactic transplantation of NSCs or ANSCs into the hippocampus. (B), Body weight (BW) was measured weekly in rats from each group (n = 6). P values were determined using Welch's t ‐tests. (C), Spleen coefficient of rats from each group at the experimental endpoint (day 28, n = 6). P values were determined using Welch's t ‐tests. (D), Sucrose preference test (SPT) assessing sucrose consumption in rats from each group before and after PBS, NSCs, or ANSCs injection (n = 6). P values were determined using Welch's t ‐tests. (E), Open field test (OFT) and elevated plus maze (EPM) behaviors of rats were recorded before and after PBS, NSCs, or ANSCs injection and analyzed using an automated animal behavior analysis system. (F), Total distance traveled during the OFT in each group. P values were determined using Welch's t ‐tests. (G), Percentage of time spent in the open arms of the EPM in each group. P values were determined using Welch's t ‐tests. (H), Representative immunofluorescence micrographs of the rat hippocampus. Red: tdTomato, green: GFAP. Scale bar, 100 µm. (I), Representative immunofluorescence micrographs of the enlarged rat hippocampus. Nuclei were stained with DAPI (blue), and transplanted cells were identified by tdTomato (red). Scale bar, 50 µm. (J), The number of tdTomato + cells with identifiable nuclear was quantified in representative microscopic fields of the NSC and ANSC group of rat hippocampal sections. (K), Detection of human‐derived transcripts in rat hippocampal RNA‐seq data using human‐specific gene sequences. Open circles indicate samples without detectable human‐specific sequences, whereas filled circles indicate samples with detectable human‐derived transcript fragments. Heatmap showing normalized read counts of detected human‐specific transcript fragments in rat hippocampal RNA‐seq data. (L), Principal component analysis (PCA) of hippocampal transcriptomes from CON, Depressed, Sham, NSC, and ANSC groups. (M), Hierarchical clustering of hippocampal transcriptomes from CON, Depressed, Sham, NSC, and ANSC groups (distance metric: 1‐Spearman correlation coefficient).

Article Snippet: The primary antibodies used included: rabbit polyclonal OCT4 (Novus Biologicals, #NBP2‐15053, 1:200), rabbit polyclonal NANOG (PeproTech, #P236, 1:200), goat polyclonal SOX2 (R&D Systems, #AF2018, 1:200), rabbit monoclonal NESTIN (Boster, #PB9874, 1:200), rabbit polyclonal PAX6 (Elabscience, #E‐AB‐61653, 1:200), rabbit polyclonal N‐Cadherin (Abcam, #ab76057, 1:200), goat polyclonal Brachyury (R&D Systems, #AF2085, 1:100), goat polyclonal SOX17 (R&D Systems, #AF1924, 1:200), mouse monoclonal NeuN (CST, #93972, 1:100), mouse monoclonal TUBB3 (Bioss, #BSM‐33177M, 1:200), rabbit polyclonal GFAP (Bioss, #bs‐0199R, 1:200), and rabbit monoclonal CDX2 (Biogenex, #Cdx2‐88).

Techniques: Transplantation Assay, Injection, Immunofluorescence, Staining, Derivative Assay, RNA Sequencing

Journal: bioRxiv

Article Title: Mouse behavioral genomics identifies Creld1 as a gatekeeper of somatosensation

doi: 10.64898/2026.03.30.715210

Figure Lengend Snippet: a , Schematic illustration for identification of Creld1-interacting protein in DRGs by immunoprecipitation and mass spectrometry. b , Representative Na v currents in DRG neurons from control and sgCreld1 mice. c , Representative Na v currents recorded in DRG neurons derived from WT and cKO mice. d , Scatterplot of normalized mRNA levels of the indicated Na v isoforms expressed in WT and cKO DRGs. e , Immunofluorescent staining of either Na v 1.7 and TUBB3 from the indicated DRG sections. Scale bar, 20 μm. f , Total density of Na v 1.7 expression of DRG neurons from WT (389 cells) and cKO (252 cells). g , Immunofluorescent staining of endogenous Creld1 and Na v 1.7 in DRG neurons after 36 h in culture using the anti-Creld1, anti-Na v 1.7 and anti-TUBB3 antibody. Scale bars, 100 μm (top); 5 μm (bottom). h , Pearson’s co-localization efficiency analysis for Na v 1.7 and Creld1 located in DRG neurons after 36 h in culture. The white circle illustrates the region of interest (ROI) used for analyzing the immunofluorescence staining of Na v 1.7 and Creld1. Each dot represents an individual cell (17 cells in total).

Article Snippet: Primary antibodies: rabbit anti-GFP (ThermoFisher Scientific, A11122, 1:500 dilution), chicken anti-GFP (Aves Labs, GFP-1020, 1:500 dilution), chicken anti-NFH (Aves Labs, NFH, 1:500 dilution), biotin IB4 (ThermoFisher Scientific, I21414, 1:100 dilution), rabbit anti-CGRP (immunostar, 24112, 1:500 dilution), mouse anti-Piezo2 (generated in our lab, 1:500 dilution), mouse anti-Trpv1 (Abcam, ab203103,1:500 dilution), human anti-Na v 1.7 (US11643458B2, a gift from Dr. Juanjuan Du at Tsinghua university, 1:500 dilution), goat anti-Creld1 (R&D systems, AF4116, 1:500 dilution), rabbit anti-TUBB3 (CST, 5666S, 1:500 dilution), rabbit anti-PGP9.5 (Cell signaling technology, 13179).

Techniques: Immunoprecipitation, Mass Spectrometry, Control, Derivative Assay, Staining, Expressing, Immunofluorescence

Journal: bioRxiv

Article Title: Human genetic variation shapes the response of neurons to interferons

doi: 10.1101/2025.05.28.653507

Figure Lengend Snippet: a ) SNPs within 500kb of the lead CASP7 eQTL are shown colored by R 2 value with respect to the lead CASP7 eQTL (green). b ) Top, example western blot for CASP7 with TUBB3 as loading control. Bottom, protein quantification normalized to TUBB3 expression (n=3 independent pairs) *p<0.01 by student’s t-test. c ) STAT1 binding at the CASP7 promoter in cells carrying two copies of the major (CTT;CTT) or minor (C;C) allele. Signal is normalized to total number of reads. d ) Relative STAT1 Cut&Tag read density across CASP7 eQTL in cells heterozygous at the allele (CTT; C) (n=4). **p<0.0001 by chi-squared test (for each experiment). H3K4me3 read density across the same site as STAT1 binding, shown as control (n=2; p-value for each chi-squared test = 0.04 and 0.52).

Article Snippet: Samples were run on NuPage Bis-Tris 4-12% gels (Thermofisher), transferred using iBlot2 (Thermofisher), and membranes were probed with anti-TUBB3 antibody (1:1000, Cell Signaling Technologies, #5568) or anti-CASP7 antibody (1:1000, Abcam, #AB32522).

Techniques: Western Blot, Control, Expressing, Binding Assay