mouse anti tgf β1 ab  (R&D Systems)

Journal: Frontiers in Immunology

Article Title: Extramedullary hematopoiesis contributes to enhanced erythropoiesis during pregnancy via TGF-β signaling

doi: 10.3389/fimmu.2023.1295717

Figure Lengend Snippet: TGF-β promoted the generation of CECs from CD34 + HSPCs. Concentrations of TGF-β in the serum of non-pregnant and pregnant women (A) and mice (B) were presented. (C) Protein expression of splenic TGF-β in non-pregnant or pregnant mice was shown. Representative flow cytometric scatter plots of HSPCs-derived CECs treated with vehicle (present as Control group) or different concentrations of TGF-β on day 7 (D) and day 11 (E) were presented. Graphical summaries of the percentages of CECs from UBMC-derived HSPCs with vehicle or different concentrations of TGF-β on day 7 (F) and day 11 (G) were shown. Statistical analysis was performed using Student’s- t test or one-way ANOVA. The results were expressed as mean ± SD. # P < 0.05 and * P < 0.01 vs. the control group. Non-P, non-pregnant women or mice; P, pregnant women or mice.

Article Snippet: In some experiments, TGF-β pathway inhibitors, including 1 μM SB431542 (a TGF-β receptor inhibitor; MedChemExpress), 1 μM Galunisertib (a TGF-β receptor inhibitor; MedChemExpress), 1 μM ITD-1 (a Smad2/3 inhibitor; MedChemExpress), and 10 μg/mL anti-mouse TGF-β Ab (Bioxcell, Lebanon, NH, USA), were added to the co-culture systems.

Techniques: Expressing, Derivative Assay

Journal: Frontiers in Immunology

Article Title: Extramedullary hematopoiesis contributes to enhanced erythropoiesis during pregnancy via TGF-β signaling

doi: 10.3389/fimmu.2023.1295717

Figure Lengend Snippet: Splenic stromal cells promoted the production of CECs from LSK cells through the TGF-β pathway. (A) Representative morphologies of splenic stromal cells and the co-culture of bone marrow LSK cells and splenic stromal cells were shown. Scale Bar: 200 μm. Flow cytometric analysis of the percentages of CD45 - splenic stromal cells (B) and the production of intracellular TGF-β in splenic stromal cells (C) of non-pregnant or pregnant mice were presented. (D) The relative protein expression of splenic TGF-β in non-pregnant and pregnant mice assessed using Western blot analysis. (E) Flow cytometric scatter plots and graphical summaries were presented to illustrate the generation of CECs from co-cultures of LSK cells and splenic stromal cells, with or without different inhibitors of the TGF-β pathway. (F) The percentages of CECs in co-cultures of LSK cells and splenic stromal cells, with or without various inhibitors of TGF-β pathway, were displayed. Statistical analysis was performed using Student’s t -test or one-way ANOVA. The results were expressed as mean ± SD. # P < 0.05 and * P < 0.01 vs. the co-culture group. Non-P, non-pregnant mice; P, pregnant mice.

Article Snippet: In some experiments, TGF-β pathway inhibitors, including 1 μM SB431542 (a TGF-β receptor inhibitor; MedChemExpress), 1 μM Galunisertib (a TGF-β receptor inhibitor; MedChemExpress), 1 μM ITD-1 (a Smad2/3 inhibitor; MedChemExpress), and 10 μg/mL anti-mouse TGF-β Ab (Bioxcell, Lebanon, NH, USA), were added to the co-culture systems.

Techniques: Co-Culture Assay, Expressing, Western Blot

Journal: Frontiers in Immunology

Article Title: Extramedullary hematopoiesis contributes to enhanced erythropoiesis during pregnancy via TGF-β signaling

doi: 10.3389/fimmu.2023.1295717

Figure Lengend Snippet: Role of splenic macrophages in the generation of CECs from LSK cells. (A) Representative flow cytometric scatter plots and a graphical summary of the percentages of splenic F4/80 + macrophages in non-pregnant and pregnant mice were shown. (B) Representative flow cytometric scatter plots and a graphical summary of the intracellular TGF-β production in splenic macrophages of non-pregnant or pregnant mice were presented. Representative flow cytometric scatter plots (C) and graphical summaries (D) of CECs generated from co-cultures of LSK cells and splenic macrophages with or without various TGF-β pathway inhibitors were shown. Statistical analysis was performed using Student’s t -test or one-way ANOVA. The results were expressed as mean ± SD. # P < 0.05 and * P < 0.01 vs. the co-culture group. Non-P, non-pregnant mice; P, pregnant mice.

Article Snippet: In some experiments, TGF-β pathway inhibitors, including 1 μM SB431542 (a TGF-β receptor inhibitor; MedChemExpress), 1 μM Galunisertib (a TGF-β receptor inhibitor; MedChemExpress), 1 μM ITD-1 (a Smad2/3 inhibitor; MedChemExpress), and 10 μg/mL anti-mouse TGF-β Ab (Bioxcell, Lebanon, NH, USA), were added to the co-culture systems.

Techniques: Generated, Co-Culture Assay

Journal: Frontiers in Immunology

Article Title: Extramedullary hematopoiesis contributes to enhanced erythropoiesis during pregnancy via TGF-β signaling

doi: 10.3389/fimmu.2023.1295717

Figure Lengend Snippet:

Article Snippet: In some experiments, TGF-β pathway inhibitors, including 1 μM SB431542 (a TGF-β receptor inhibitor; MedChemExpress), 1 μM Galunisertib (a TGF-β receptor inhibitor; MedChemExpress), 1 μM ITD-1 (a Smad2/3 inhibitor; MedChemExpress), and 10 μg/mL anti-mouse TGF-β Ab (Bioxcell, Lebanon, NH, USA), were added to the co-culture systems.

Techniques: Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Nitric Oxide Production by Myeloid Derived Suppressor Cells Plays a Role in Impairing Fc Receptor-Mediated Natural Killer Cell Function.

doi: 10.1158/1078-0432.CCR-17-0691

Figure Lengend Snippet: Female Balb/c mice were inoculated with 1x105 4T1 tumor cells or were left un-injected (no tumor-control). Following the establishment of tumors, mice were treated daily with (a) intraperitoneal injections of IgG (250 μg), anti-TGF-β antibody (200 μg), or anti-IL-10 antibody (250 μg) for four consecutive days prior to NK cell isolation for the ADCC assay. The mice were sacrificed 24 hrs after the last treatment. Each group consists of pooled samples from spleens of 4-5 mice. Values represent mean ± SD from one experiment. (b) Mice were treated with PBS (vehicle), arginase inhibitor nor-NOHA (20 mg/kg) i.p or the IDO inhibitor 3-methyltryptophan (MT, 400 mg/kg) via oral gavage prior to NK cell isolation for the ADCC assay. (c) Mice were given intraperitoneal injections of PBS (vehicle) or iNOS inhibitor, L-NIL (20 mg/kg) for one week. NK cells isolated from the spleen were employed in a standard ADCC assay using trastuzumab-coated CT26-HER2 positive tumor cells as targets. (d) NK cells and MDSC from the peripheral blood of melanoma patients were co-cultured overnight at a 1:1 ratio with or without the nitric oxide inhibitor L-NIL (2.5 mM). ADCC function of NK cells displayed as the mean percent lysis of cetuximab-coated HT-29 tumor cells at the 10:1 effector to target ratio. Means ± SE from four independent experiments are shown, p<0.05. Significance was determined using a linear mixed model. Treatment of NK cells with L-NIL alone did not enhance ADCC activity (not shown). (e) Autologous NK cells and MDSC isolated from peripheral blood of melanoma patients were co-cultured in 96 well plates coated with human IgG or media with or without the iNOS inhibitor L-NIL (2.5 mM). Supernatants were harvested after 48 hrs and analyzed for levels of IFNγ by ELISA. Values represent mean ± SE from three independent experiments, p<0.05. Significance was determined using a linear mixed model. (f) p-Erk expression in NK cells co-cultured overnight with MDSC in the presence or absence of L-NIL (2.5 mM). p-Erk levels are expressed as the average fold change ± SE from three independent experiments, p<0.05. Significance was determined using a linear mixed model.

Article Snippet: To inhibit MDSC function Balb/c mice received intraperitoneal injections of the following agents for 5 consecutive days: L-NIL (20 mg/kg), anti-TGF-β Ab (200 μg, BioXcell, clone 1D11.16.8) ( 25 ), anti-IL-10 Ab (250 μg, BioXcell, clone JES5-2A5), the arginase inhibitor nor-NOHA (20 mg/kg) ( 27 ) or the IDO inhibitor, 1-Methyl-D-tryptophan via oral gavage (400 mg/kg, Sigma Aldrich) ( 26 ).

Techniques: Injection, Cell Isolation, ADCC Assay, Isolation, Cell Culture, Lysis, Activity Assay, Enzyme-linked Immunosorbent Assay, Expressing