tgf-β monoclonal mouse neutralizing antibody tgfmab clone 1d11  (Bio X Cell)

Journal: Frontiers in Cell and Developmental Biology

Article Title: Overexpression of MUC1 Induces Non-Canonical TGF-β Signaling in Pancreatic Ductal Adenocarcinoma

doi: 10.3389/fcell.2022.821875

Figure Lengend Snippet: Heatmap showing top 30 differentially expressed genes in high/moderate vs low MUC1 PDA samples from TCGA. (A) Top panel shows the color key for MUC1 expression in the 29 PDA samples. Right hand side shows the color key histogram for expression levels of each gene named on the right. Left hand side color key shows the genes associated with each of the three pathways in pink (TGF-β), green (MAPK) and peach (BMP). Genes with a false discovery rate adjusted p < 0.05 are shown. (B) Kaplan-Meier curve for overall survival (OS) in the 29 PDA patients from TCGA in low (blue) vs high/moderate (red) groups are shown.

Article Snippet: Groups 2 and 4 were treated with the monoclonal TGF-β neutralizing antibody (LifeTech) (20ug/100ul per mouse) three times a week for 2 weeks.

Techniques: Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Overexpression of MUC1 Induces Non-Canonical TGF-β Signaling in Pancreatic Ductal Adenocarcinoma

doi: 10.3389/fcell.2022.821875

Figure Lengend Snippet: Overexpression of MUC1 leads to increased phosphorylation of JNK and c-Myc and knockdown of MUC1 reduces phosphorylation of JNK and c-Myc. (A) Western blot expression of phosphorylation of JNK and c-Myc compared to total JNK and total c-Myc in MiaPaca2 vs MiaPaca2. MUC1 cells in response to 10 ng/ml of TGF-β at 10 min. (B) Western blot expression of phosphorylation of JNK and c-Myc compared to total JNK and total c-Myc in HPAFII cells treated with control siRNA vs MUC1 siRNA in response to 10 ng/ml of TGF-β at 10 min. (C) Densitometric analysis of fold change of expressions of pJNK/Total JNK and p-c-Myc/Total c-Myc normalized to endogenous β-actin is presented in MiaPaca2 cells. (D) Densitometric analysis of fold change of expressions of pJNK/Total JNK and p-c-Myc/Total c-Myc normalized to endogenous β-actin is presented in HPAFII cells. (E) Knockdown efficiency of MUC1 in HPAFII after 72 h of siRNA treatment. Data are presented as means ± SEM of n = 3; Unpaired Student’s t-test and one-way ANOVA were used to analyze the differences between treatment groups. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Groups 2 and 4 were treated with the monoclonal TGF-β neutralizing antibody (LifeTech) (20ug/100ul per mouse) three times a week for 2 weeks.

Techniques: Over Expression, Western Blot, Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Overexpression of MUC1 Induces Non-Canonical TGF-β Signaling in Pancreatic Ductal Adenocarcinoma

doi: 10.3389/fcell.2022.821875

Figure Lengend Snippet: TGF-β exposure increases viability in cells with high MUC1 and reduces viability in low MUC1 PDA cells. MTT cell viability assay on (A) MiaPaca2.Neo cells with 10 ng/ml of TGF-β for 48 h. (B) HPAFII and (C) MiaPaca2. MUC1 cells with 10 ng/ml of TGF-β for 72 h. (D) HPAFII treated with control or MUC1 siRNA for 72 h followed by treatment with 10ng/ml of TGF-β for 24 h. All data are shown as means ± SEM of n = 3. Unpaired t-test was performed to compare between treated and untreated cells for each one of experiments A-C and two-way ANOVA was used to compare between untreated and treated in HPAFII.controlsiRNA and HPAFII.MUC1siRNA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Groups 2 and 4 were treated with the monoclonal TGF-β neutralizing antibody (LifeTech) (20ug/100ul per mouse) three times a week for 2 weeks.

Techniques: Viability Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Overexpression of MUC1 Induces Non-Canonical TGF-β Signaling in Pancreatic Ductal Adenocarcinoma

doi: 10.3389/fcell.2022.821875

Figure Lengend Snippet: TGF-β neutralizing antibody treatment significantly reduced high-MUC1 (HPAFII) but not low MUC1 (MiaPaca2) tumor growth in vivo . (A) A schematic of the xenograft study showing the treatment with control IgG and anti-TGF-β antibody (20 ug/100 ul per mouse). (B) On the left: Tumor growth of HPAFII (n = 5 for TGF-β neutralizing Ab and n = 4 for IgG isotype) is shown. On the right: Tumor growth of MiaPaca2 (n = 6 for both groups) is shown. Tumor growth was determined biweekly by caliper measurements and tumor size in mm 3 is plotted. (C) Wet weight of HPAFII tumors (left) and MiaPaca2 tumors (right) respectively are shown. Two-way ANOVA was used to compare between the different treatment groups. * p <0.05, NS: non-significant. (D) Immunohistochemistry showing expression of MUC1 in MiaPaca2 (left) and HPAFII (right) tumors.

Article Snippet: Groups 2 and 4 were treated with the monoclonal TGF-β neutralizing antibody (LifeTech) (20ug/100ul per mouse) three times a week for 2 weeks.

Techniques: In Vivo, Immunohistochemistry, Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Overexpression of MUC1 Induces Non-Canonical TGF-β Signaling in Pancreatic Ductal Adenocarcinoma

doi: 10.3389/fcell.2022.821875

Figure Lengend Snippet: Schematic diagram of the proposed mechanism of TGF-β signaling and functions in high versus low MUC1 PDA . Left panel shows activation of SMAD-dependent canonical pathway in low-MUC1 PDA cells. TGF-β ligands bind to the membranous TGF-β receptor (TGF-βRII) homodimers with high affinity. TGF-βRII binding allows dimerization with TGF-β type I receptor (TGF-βRI) homodimers, activation of the TGF-βRI kinase domain and signal transduction via phosphorylation of the C-terminus of receptor-regulated SMADs (R-SMAD), SMAD2 and SMAD3. The SMAD2/3 dimer then forms a heterotrimeric complex with SMAD4 which translocates in the nucleus ( ; ). This leads to growth inhibition, cell cycle arrest and apoptosis of PDA cells, thus TGF-β acts as a tumor suppressor. Right panel shows activation of SMAD-independent non-canonical pathway in high-MUC1 PDA cells. In this pathway, binding of TGF-β mainly to TGF-β-RII most likely increases phosphorylation of c-SRC which in turn phosphorylates MAPK, followed by JNK and c-Myc . This phosphorylation cascade activates the MAPK/JNK pathway and stabilizes c-Myc which translocates into the nucleus to increase transcription of oncogenic proteins and leads to increased growth, invasion and EMT of PDA cells . MUC1-CT also aids in the process by its oncogenic signaling. Thus, in high-MUC1 PDA cells TGF-β acts as a pro-tumorigenic cytokine. The schematic was created with BioRender.com .

Article Snippet: Groups 2 and 4 were treated with the monoclonal TGF-β neutralizing antibody (LifeTech) (20ug/100ul per mouse) three times a week for 2 weeks.

Techniques: Activation Assay, Binding Assay, Transduction, Inhibition

Journal: Experimental & molecular medicine

Article Title: Insulin resistance mediates high-fat diet-induced pulmonary fibrosis and airway hyperresponsiveness through the TGF-β1 pathway.

doi: 10.1038/s12276-019-0258-7

Figure Lengend Snippet: Fig. 3 AHR and inflammatory features of naive HFD and OVA-HFD models. AHR to MCh (a), inflammatory cell profile in the BALF (b), IL-4 (c) and IL-5 levels (d), and TGF-β1 (e) concentrations in lung homogenates. Immunohistochemical staining for TGF-β1 (F) in tissue from naïve HFD, OVA-HFD, naive ND, and OVA-ND mice are shown. †P < 0.05; ‡P < 0.01; *P < 0.001. AHR airway hyperresponsiveness, MCh methacholine, BALF bronchoalveolar lavage fluid, HFD high-fat-diet, IL interleukin, OVA ovalbumin, ND normal diet, TGF transforming growth factor

Article Snippet: To block TGF-β1, a rabbit anti-mouse TGF-β neutralizing mAb (100 μg/mouse; R&D Systems Inc., Minneapolis, MN, USA) or rabbit IgG antibody (100 μg/ mouse; R&D Systems) was administered once via the intravenous (i.v.) route 4 h before the first OVA challenge (Fig. 1).

Techniques: Immunohistochemical staining, Staining