mouse monoclonal anti h4  (Cell Signaling Technology Inc)

Journal: Frontiers in Immunology

Article Title: Peripheral blood T-cell subsets combined with EGRIS score predict the need for mechanical ventilation in Guillain–Barré syndrome

doi: 10.3389/fimmu.2026.1747416

Figure Lengend Snippet: Association of peripheral blood lymphocyte counts with the need for mechanical ventilation in patients with Guillain–Barré syndrome (GBS). (A) ROC analysis of CD8 + T cell and CD4 + T cell counts and the predictive value of Erasmus GBS Respiratory Insufficiency Score (EGRIS) for mechanical ventilation in GBS. P < 0.05 was considered significant. (B) Multivariate logistic regression analysis of gender, age, CD8 + T cell counts and CD4 + T cell counts and EGRIS in GBS patients requiring mechanical ventilation. P < 0.05 was considered significant.

Article Snippet: Sections were incubated with mouse anti-CD4 monoclonal antibody (Protein tech) overnight at 4°C and then with FITC-conjugated goat anti-mouse IgG (H + L) (Boster Biological Technology) for 1 hour at room temperature.

Techniques:

Journal: Frontiers in Immunology

Article Title: Peripheral blood T-cell subsets combined with EGRIS score predict the need for mechanical ventilation in Guillain–Barré syndrome

doi: 10.3389/fimmu.2026.1747416

Figure Lengend Snippet: CD4 + T cell expression in the sciatic nerve of experimental autoimmune neuritis (EAN) rats, n = 4. (A) Immunofluorescence staining of rat sciatic nerve tissue. (original magnification, 40×). Green fluorescence indicates CD4 + T cells, blue fluorescence indicates DAPI, and the cells outlined in white and green represent CD4 + T cells. (B) Preparation of EAN rats, neurological function monitoring, and tissue collection timeline. (C) Neurological function score curve of EAN rats within 30 days after immunization. (D) Cell counts are expressed as mean ± standard deviation (SD). One-way ANOVA was used to assess the number of CD4 + T cells in the sciatic nerves of EAN rats at different disease stages, followed by Tukey’s post hoc test for multiple group comparisons. A p-value < 0.05 was considered statistically significant. Statistical differences between groups are indicated by asterisks: * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Sections were incubated with mouse anti-CD4 monoclonal antibody (Protein tech) overnight at 4°C and then with FITC-conjugated goat anti-mouse IgG (H + L) (Boster Biological Technology) for 1 hour at room temperature.

Techniques: Expressing, Immunofluorescence, Staining, Fluorescence, Standard Deviation

Journal: bioRxiv

Article Title: EpiBrain: the brain’s epigenetic landscape in a snapshot

doi: 10.64898/2026.01.22.701139

Figure Lengend Snippet: ( A ) A schematic illustration of the EpiBrain imaging and signal detection approach. Larval zebrafish are counterstained for histone modification (HM) and histone H3 (H3) or DNA modification (DM) and DNA. Whole-brain images taken from control and treatment groups are shown with differential levels of abundance for HM and DM in distinct brain regions. Images are registered to a reference brain for signal detection. Brain regions with elevated HM/DM in the treatment group as compared to the control group are labeled in green, whereas regions with reduced HM/DM in the treatment group are labeled in magenta. ( B ) Schematic illustrations of three EpiBrain staining protocols. All three protocols were developed based on a base protocol which consists of the following steps: fixation (FX), bleaching (BL), antigen retrieval (AR), permeabilization (PM), blocking (BK), primary antibody incubation (1 st Ab), and secondary antibody incubation (2 nd AB). The EpiBrain-H3(K27/K36) protocol counterstains modifications on the H3 tail that are close to the H3 core (i.e., lysine residuals K27 and K36) against H3. H3 is detected using an anti-H3-tail antibody (H3Tail) that targets the N-terminus of H3. Different from the base protocol, DIMEB is added during primary and secondary antibody incubations to boost antibody penetration. A longer incubation time (↑IT) is also applied to improve antibody binding. The EpiBrain-DNA protocol counterstains DNA modifications (i.e., 5mC and 5hmC) against the DNA. Several additional treatment steps are added between the FX and BL steps of the base protocol, including hydrogel polymerization (HP), SDS treatment, and a post-fixation step (2 nd FX). The AR and PM steps in the base protocol are replaced by a HCl-based antigen retrieval step (AR: HCl). Similar to EpiBrain-H3(K27/K36), DIMEB and a longer incubation time are applied to boost antibody penetration. In addition, samples are centrifuged and incubated at 37°C (CF) during primary and secondary antibody incubations to further improve antibody staining. Propidium Iodide (PI) is applied after antibody incubations to label DNA. The EpiBrain-H3(K4/K9) protocol counterstains modifications on the N-terminus of the histone H3 tail (i.e., lysine residuals K4 and K9) against H3. H3 is detected using an anti-H3-core antibody targeting the C-terminus of H3. This protocol is otherwise similar to the EpiBrain-DNA protocol minus the PI staining step. ( C-G ) EpiBrain-H3(K27/K36) staining and imaging analysis detected expected changes in the zebrafish brain’s histone modification state following targeted perturbations. The level of H3K27me3 in the brain is globally reduced by the Ezh2 inhibitor UNC1999 (C) and elevated by the H3K27me3-demethylase inhibitor GSK-J4 (D). H3K27ac level is reduced by the histone acetyltransferase p300 inhibitor C646 (E) and increased by the HDAC inhibitor VPA (F). H3K36me3 level is reduced by knocking out (KO) the histone methyltransferase setd2 (G). Each panel consist of three images. The two images on the left are dorsal and lateral views of the brain, with changes in the relative abundance of an epigenetic modification shown in colors: green represents elevation compared to the control, and magenta represents reduction. A: anterior. P Posterior. L: left. R: right. V: ventral. D: dorsal. The image on the right highlights brain regions with significant changes in the epigenetic modification. These brain regions are labeled in colors, with green representing regions with elevated levels of epigenetic modification compared to the control and magenta representing reduced levels of modification. The brightness of the color correlates with the degree of significance. ( H-K ) EpiBrain-DNA staining and imaging analysis detects expected changes in the zebrafish brain’s DNA methylation state following targeted perturbations. The level of 5hmC is reduced by knocking out the 5mC dioxygenases tet1 (H), tet2 (I), and tet3 (J). The level of 5mC is reduced by the DNMT inhibitor decitabine (K). ( L-O ) EpiBrain-H3(K4/K9) staining and imaging analysis detects expected changes in the zebrafish brain’s histone modification state following targeted perturbations. H3K4me1 level is reduced by knocking out the lysine methyltransferase setd7 (L). H3K4me3 level is also largely reduced by setd7 knockout, but with an elevation in the dorsal medial region (M). H3K9me2 level is reduced by the lysine methyltransferases G9a and GLP inhibitor UNC0642 (N). H3K9ac level is reduced by C646 (O). Brain regions are identified based on alignment with the zBrain atlas following brain registration. The top differentially regulated brain regions are labeled in the images. Raw analysis results for all differentially regulated brain regions and their signal intensities in images shown in are summarized in Supplementary Data 1. AP: area postrema. GAD1BS1: gad1b stripe 1 (spinal cord). GAD1BC14: gad1b cluster 14. GLYT2S1: glyt2 stripe 1. HCRTRS4: 6.7FDhcrtR-Gal4 stripe 4. LCC: locus caudalis cerebelli. NA: noradrendergic neurons of the interfascicular and vagal areas. OBDN: olfactory bulb dopaminergic neuron. PG: pineal gland. PFT1A: pft1a stripe. QRFP: qrfp neuron cluster sparse. S1181t: s1181t cluster. SPDC: subpallial dopaminergic cluster. TH: small cluster of TH-stained neurons. TL: torus longitudinalis. VGLUT2S1: vglut2 stripe 1 (rhombencephalon). VMAT2: vmat2 cluster (telencephalon). VMAT2S1: vmat2 stripe1 (rhombencephalon).

Article Snippet: A mouse monoclonal antibody against histone H3 was purchased from Proteintech (CAT#68345).

Techniques: Imaging, Modification, Control, Labeling, Staining, Blocking Assay, Incubation, Binding Assay, DNA Methylation Assay, Knock-Out, Olfactory

Journal: Andrology

Article Title: The Oxytocin Receptor in Spermatozoa May Originate From Both Spermatogenesis and Epididymal Maturation, and Regulates Capacitation

doi: 10.1111/andr.70123

Figure Lengend Snippet: Confocal microscopy (immunofluorescence) for pig testis and epididymis, and transmission electron microscopy (TEM) for pig epididymis. (A) 3D projection (z‐stack) in testis. Immunolabeling for the oxytocin receptor (OR) was observed in germinal cells (arrow), whereas while myoid cells were negative (arrowhead). Insert detail: The OR was found in the post‐acrosomal region of pig spermatozoa in the lumen of seminiferous epithelium (arrow). (B) Spermatozoa from the corpus of the epididymis. Strong labeling for the OR was observed in the apical blebs, which showed vesicular heterogeneity in content and size (arrow). (C) 3D projection (z‐stack) of spermatozoa in the corpus epididymis. Labeling for the OR was observed in the post‐acrosomal region of spermatozoa (arrow) and probably in the disintegrated apical blebs in close contact with spermatozoa (arrowhead). Insert detail: Positivity for the OR in the sperm tail. (D–I). Colocalization of CD81, caveolin‐1, and the OR in the epididymal tissue. (D) The apical blebs (arrow) and supranuclear region of the epididymal principal cells (star) were positive for tetraspanin CD81. (E) OR positivity was observed in the apical blebs (arrow), principal cells (star) and smooth muscle cell layer (arrowhead). (F) Staining with SYTOX orange, which stains the nuclei of spermatozoa in the lumen (star), the nuclei of the cells in the epididymal epithelium (arrow) and the nuclei of smooth muscle cells in the muscular wall (arrowhead). (G) Caveolin‐1 immunostaining in apical blebs (arrow), cells of the epididymal epithelium (star), and muscle layer (arrowhead). (H) Merge of the above four channels, which showed the colocalization of the OR, CD81, and caveolin‐1 in the apical blebs (arrows) of the epididymal epithelium. The supranuclear region of the principal cells also depicted strong colocalization of these three antibodies (star). (I) Merge of the four channels of the image (B), showing in more detail the colocalization of CD81, cav‐1, and OR in the apical blebs and derived structures (arrows). (J) 3D projection (z‐stack) of spermatozoa from cauda epididymis. Immunolabeling for the OR is detected in the sperm tail (white arrow). (K) A merged four‐channel image shows an apical bleb emerging from the epididymal epithelium in the caput region (white arrow), along with free apical blebs in the lumen in proximity to spermatozoa (arrowhead). Insert detail: higher magnification of the free apical blebs (arrowhead) shows the presence of the OR within their lumen. (L–O) Transmission electron microscopy (TEM). A potential sequence of apical bleb disintegration into smaller vesicular substructures is shown. (L) An apical bleb emerging from an epididymal epithelial cell (corpus) containing heterogeneous multivesicular content. Note the parallelism with confocal microscopy images. (M) Free apical bleb containing heterogeneous substructures (star) in the epididymal lumen. (N) Putative free vesicular substructures resembling those found inside the apical bleb shown in M (star). (O) Smaller individualized vesicular structures, likely exosomes (arrow), exhibiting diverse morphologies and localized among or adjacent to spermatozoa. Free vesicular substructures (star) appear to release these exosomes (arrowhead). Other single extracellular vesicles larger (red arrow) than exosomes are also observed both in the epididymal lumen and within the substructures (red arrowhead) of the apical blebs (star). Scale bars: (A–C, I, J) 10 µm; (D–H, K) 20 µm; (L) 5 µm; (M, N) 2 µm; (O) 1 µm. Inserts detail 10 µm.

Article Snippet: Samples were incubated with three antibodies; a monoclonal mouse antibody against tetraspanin CD81 (Proteintech, 66866‐1, Manchester, United Kingdom) at a dilution of 1/50 (v/v), a polyclonal goat antibody against caveolin‐1 at a dilution of 1/100 (v/v) (Abcam, ab36152, Cambridge, United Kingdom), and a polyclonal rabbit antibody against OR at a dilution 1/50 (v/v) (Proteintech, 23045‐1‐AP), in PBS containing 1% BSA, at 4°C overnight in a humidity chamber.

Techniques: Confocal Microscopy, Immunofluorescence, Transmission Assay, Electron Microscopy, Immunolabeling, Labeling, Staining, Immunostaining, Derivative Assay, Sequencing

Journal: Cell reports

Article Title: Poxvirus A51R proteins regulate microtubule stability and antagonize a cell-intrinsic antiviral response

doi: 10.1016/j.celrep.2024.113882

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-Histone H4 (clone F-9) , Santa Cruz Biotechnology , Cat# sc-25260; RRID: AB_2118623.

Techniques: Virus, Control, Recombinant, Lysis, Staining, Protease Inhibitor, Magnetic Beads, Modification, Saline, Expressing, Transfection, Blocking Assay, Fractionation, HTS Assay, Plasmid Preparation, Software, Imaging