Mouse TGF-beta 2 Antibody

Detection of Human and Mouse TGF‑ beta 2 by Western Blot. Western blot shows lysates of Y-79 human retinoblastoma cell line, HUVEC human umbilical vein endothelial cells, NIH-3T3 mouse embryonic fibroblast cell line, 4T1 mouse breast cancer cell line, and NMuMG mouse mammary gland epithelial cell line. PVDF membrane was probed with 2 µg/mL of Rat Anti-Mouse TGF-beta 2 Monoclonal Antibody (Catalog # MAB73461) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for TGF-beta 2 at approximately 52 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

TGF‑ beta 2 in Mouse Embryo. TGF-beta 2 was detected in immersion fixed frozen sections of mouse embryo (15 d.p.c.) using Rat Anti-Mouse TGF-beta 2 Monoclonal Antibody (Catalog # MAB73461) at 25 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS017) and counterstained with hematoxylin (blue). Specific staining was localized to neuronal processes. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Detection of Human TGF-beta 2 by Western Blot LOXL2 acts with TGF-beta through PI3K/ATK/mTORC1 signalling to control myofibroblast transformation and migration.(a) Western blot analysis of LOXL2, SMAD2, p-SMAD2 and GAPDH in human primary cardiac fibroblasts transfected with control (Ctrl) or LOXL2 siRNA. (b) Quantification of TGF-beta isoforms in the culture media of human primary cardiac fibroblasts transfected with control or LOXL2 siRNA (n=4). P value: Student's t-test. Error bar: s.e.m. (c) Changes of COLA1, alpha -SMA and FN1 mRNA in human primary cardiac fibroblasts transfected with control (Ctrl) or LOXL2 siRNA. (d) Western blot of LOXL2, p-AKT, AKT, p-S6K, S6K and p-4E-BP1 with or without PI3K inhibitors LY294002/PI828 (10 μM each), PI3K alpha inhibitors A66/BYL719 and mTORC1 inhibitor Rapamycine (0.1 μM) in cells infected with Ad_GFP/Ad_LOXL2. (e) TGF-beta 2 protein in the culture media (n=4). P value: Student's t-test. Error bar: s.e.m. (f,g) Western blot (f) and quantification (g) of TGF-beta 2 protein in the mice heart ventricles 6 weeks after sham/TAC operation. n=4 mice per group. P value: Student's t-test. Error bar: s.e.m. (h) A signalling cascade from LOXL2 to PI3K/AKT/mTORC1 to TGF-beta 2 translation. (i) Representative phase-contrast image of human primary cardiac fibroblasts 72 h after transfection with control (Ctrl) or LOXL2 siRNA. Scale bars, 50 μm. (j) Immunostaining of Col1A in IgG1- or alpha -LOXL2-treated mice 10 weeks (10W) after sham or TAC operation. Scale bars, 100 μm. Blue: haematoxylin. Brown: Col1A. (k,l) Gap closure assay of fibroblast migration in control, TGF-beta 2, alpha -LOXL2 (k) groups and quantification of cells migrating into the gap (l). Scale bars, 200 μm. n=5–10, P value: Student's t-test. Error bar: s.e.m. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27966531), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse TGF-beta 2 by Western Blot MicroRNA (miRNA) 466a-3p transfection inhibits regulatory T cell (Treg) polarization. Purified naïve CD4+ T cells were cultured under Treg-polarizing conditions along with the indicated mimic, control, or inhibitor conditions. Cells were harvested 48 h after addition of cytokines and miRNA mimics, inhibitors, or controls and subject to flow cytometry, immunoblot and quantitative real-time-PCR. The success of Treg polarization is examined as (A) representative dot plots gated on CD25HI cells and quantified in (B,C). Representative immunoblots of indicated proteins are presented in (D,F), along with associated densitometric measurements of transforming growth factor-beta 2 (TGF-beta 2) and TGF-beta R3 (E), and quantification of activated Smad 2, 3, and 4 (G). CD4+ cells were purified from naïve mouse lymph nodes and stimulated ex vivo with CD3 (3 µg/mL) and CD28 (3 µg/mL) for 48 h and administered Locked Nucleic Acid or controls at the time of seeding. Quantification of flow cytometry data from LAP-expressing FoxP3 positive Treg cells. (H) Purified naïve CD4+ T cells were cultured with either TGF-beta 1 (5 ng/mL) or TGF-beta 2 (5 ng/mL), along with CD3 (3 µg/mL), CD28 (3 µg/mL), and IL-2 (5 ng/mL) for 5 days. (I) representative dot plots of FoxP3, CD4-positive Tregs gated on CD25HI, (J), and their associated CD278 (ICOS) expression. Data are presented as mean ± SEM of three independent transfection experiments. *P < 0.05, **P < 0.005, ****P < 0.0001 by ANOVA with Tukey’s multiple comparison test. Image collected and cropped by CiteAb from the following publication (https://journal.frontiersin.org/article/10.3389/fimmu.2018.00688/full), licensed under a CC-BY license. Not internally tested by R&D Systems.