mouse anti ib4  (Vector Laboratories)

Journal: The Journal of Clinical Investigation

Article Title: METTL14-mediated m6A epitranscriptomic modification contributes to chemotherapy-induced neuropathic pain by stabilizing GluN2A expression via IGF2BP2

doi: 10.1172/JCI174847

Figure Lengend Snippet: ( A ) Experimental paradigm for METTL14 expression profile analysis. ( B ) Left: Representative immunofluorescence images show colabeling of METTL14 and NeuN in the rat DRG 10 days after treatment with saline or PCX. Right: The percentage of DRG neurons colabeled by METTL14 and NeuN in the L4 and L5 DRGs 10 days after treatment with saline or PCX ( n = 9 DRGs from 3 rats per group, Student’s t test). Scale bars: 100 μm. ( C ) Representative double immunofluorescence staining and quantification showing the colocalization of METTL14 with IB4, CGRP, and NF200 in rat DRG (at least 9 DRG slices from 4 rats per group). Scale bars: 100 μm. ( D ) Representative double immunofluorescence staining and quantification showing the colocalization of METTL14 with NeuN and glutamine synthetase (GS) in human DRG ( n = 10 DRG slices per group). Scale bars: 50 μm. ( E ) Representative immunofluorescence staining and quantification showing colabeling and alteration of METTL14 and DAPI in patients ( n = 3 patients per group, Student’s t test). Scale bars: 100 μm. Data are shown as the mean ± SEM. * P < 0.05, **** P < 0.0001.

Article Snippet: All sections were blocked with PBS containing 5% goat serum at 26°C for 1 hour and then incubated overnight at 4°C with the following primary antibodies: rabbit anti-METTL14 (1:200; A8530, ABclonal), mouse anti-METTL14 (1:200; CL4252, Abcam), mouse anti-NeuN (1:100; MAB377, Millipore), goat anti-GFAP (1:300; ab53554, Abcam), mouse anti-Iba1 (1:300; ab5076, Abcam), mouse anti–glutamine synthetase (1:200; MA5-27749, Invitrogen), mouse anti-CGRP (1:200; ab81887, Abcam), mouse anti-NF200 (1:200; N0142, MilliporeSigma), mouse anti-IB4 (1:50; L2895, MilliporeSigma), rabbit anti-DBP (1:200; 12662-1-AP, Proteintech), and mouse anti-FLAG (1:1,000; 66008-4-Ig, Proteintech).

Techniques: Expressing, Immunofluorescence, Saline, Double Immunofluorescence Staining, Staining

Journal: CNS Neuroscience & Therapeutics

Article Title: The G protein‐coupled estrogen receptor of the trigeminal ganglion regulates acute and chronic itch in mice

doi: 10.1111/cns.14367

Figure Lengend Snippet: Expression and distribution of the G protein‐coupled estrogen receptor (GPER) in trigeminal ganglion (TG) neurons of the mouse. (A) Schematic diagram of cholera toxin subunit B (CTB) injection in mouse and representative immunofluorescence images of CTB‐labeled TG neurons innervating the mouse cheek colocalized with GPER, along with the quantitative analysis of the overlap percentage. (B–D) Double immunofluorescence staining showing the colocalization of GPER with IB4 (B), CGRP (C) and NF200 (D), along with quantitative analysis of the overlap percentage ( n = 4 mice per group, scale bar: 100 or 40 μm).

Article Snippet: Primary antibodies used in these experiments were rabbit anti‐GPER (1:1000, Lifespan), rabbit anti‐c‐Fos (1:500, Abcam), mouse anti‐neuroflament‐200 (NF200; 1:1000, Abcam), mouse anti‐calcitonin gene‐related peptide (CGRP; 1:100, Abcam), mouse anti‐isolectin B4 conjugated to fluorescein isothiocyanate (IB4‐FITC; 1:1000, Sigma), rabbit anti‐TRPA1 (1:500, Abcam), and guinea pig anti‐TRPV1 (1:1000, Millipore).

Techniques: Expressing, Injection, Immunofluorescence, Labeling, Double Immunofluorescence Staining

Journal: CNS Neuroscience & Therapeutics

Article Title: The G protein‐coupled estrogen receptor of the trigeminal ganglion regulates acute and chronic itch in mice

doi: 10.1111/cns.14367

Figure Lengend Snippet: Gper deficiency enhances the function of TRPA1 and TRPV1 in TG neurons. (A) Representative immunofluorescence images showing the colocation of GPER with TRPA1. (B) Quantitative analysis of the percentage of GPER + /TRPA1 + neurons in total GPER + neurons of the TG ( n = 4 mice per group, scale bar: 100 or 40 μm). (C) Representative immunofluorescence images showing colocation of GPER and TRPV1. (D) Quantitative analysis of the percentage of GPER + /TRPV1 + neurons in total GPER + neurons of the TG ( n = 4 mice per group, scale bar: 100 or 40 μm). (E) Representative fluorescence images of mouse TG neurons loaded with Fura‐2 at the baseline and after application of allyl isothiocyanate (AITC, 100 μM) or capsaicin (Cap, 1 μM). Arrows indicate TG neurons responsive to the agent (scale bar: 200 μm). (F) Representative traces of Ca 2+ responses evoked by AITC or Cap in TG neurons of WT (black) and Gper −/− (red) mice. Black bars above the traces show the duration of the chemical treatment. (G–J) In comparison with WT mice, the percentage of AITC‐responsive (G) or Cap‐responsive neurons (I) was significantly increased in Gper −/− mice (** p < 0.01, *** p < 0.001, χ 2 test). In comparison with WT mice, the Ca 2+ signal amplitude induced by AITC (H) or Cap (J) was significantly higher in Gper −/− mice (** p < 0.01, *** p < 0.001, unpaired Student's t‐ test).

Article Snippet: Primary antibodies used in these experiments were rabbit anti‐GPER (1:1000, Lifespan), rabbit anti‐c‐Fos (1:500, Abcam), mouse anti‐neuroflament‐200 (NF200; 1:1000, Abcam), mouse anti‐calcitonin gene‐related peptide (CGRP; 1:100, Abcam), mouse anti‐isolectin B4 conjugated to fluorescein isothiocyanate (IB4‐FITC; 1:1000, Sigma), rabbit anti‐TRPA1 (1:500, Abcam), and guinea pig anti‐TRPV1 (1:1000, Millipore).

Techniques: Immunofluorescence, Fluorescence, Comparison