tgfβriii  (Proteintech)

Journal: Nature Immunology

Article Title: Spatial patterning of fibroblast TGFβ signaling underlies treatment resistance in rheumatoid arthritis

doi: 10.1038/s41590-025-02386-2

Figure Lengend Snippet: a – d , A fixed number of fibroblasts were seeded with varying numbers of HUVECs. Monoculture: 5,000 fibroblasts, 5:1 ratio: 5,000 fibroblasts and 1,000 HUVECs, 2:1 ratio: 5,000 fibroblasts and 2,500 HUVECs, 1:1 ratio: 5,000 fibroblasts and 5,000 HUVECs. a , b , ELISA quantification of POSTN ( a ) and COMP ( b ) production during co-culture. P values in the line graphs are displayed for the comparisons between monoculture and 1:1 ratio. The respective bar charts to the right represent the area under the curve. c , d , Flow cytometric quantification of fibroblast TGFβRII ( c ) and TGFβRIII ( d ) at the indicated days during co-culture with ECs. P values are shown for comparisons between monoculture and 1:1 ratio. Representative flow cytometry histograms and mean fluorescence intensity (MFI) quantification for day 7 of co-culture are shown. e , Gating strategy for classifying COMP hi and POSTN hi fibroblasts from the Xenium-profiled co-culture (mutually exclusive top quantile of cells expressing each gene, with POSTN ≤ 3.5 for COMP hi ). f , Representative examples of COMP hi and POSTN hi fibroblasts in 2D culture are shown with transcripts. g , Violin plot showing the distribution of TGFBR2 and TGFBR3 transcripts on POSTN hi and COMP hi gated fibroblasts in 2D co-culture, 3D co-culture and RA synovium. h , Representative example ( n = 4 RA synovial tissue) of gated (mutually exclusive top quantiles of expressing cells) COMP hi and POSTN hi synovial fibroblasts with transcripts. i , j , ELISA quantification of COMP ( i ) and POSTN ( j ) production from co-culture of fibroblasts overexpressing (OE) GFP , TGFBR2 or TGFBR3 with ECs in a 1:1 ratio. P values are shown for the comparison between TGFBR3 -OE and GFP -OE data points. The bar plot on the right represents the area under the curve. Data points are shown as the mean ± s.d., represent n = 3 biological replicates and are representative of at least two independent experiments. For statistical analysis, a two-tailed Student’s t -test was used for a – d , i and j , and a two-sided Wilcoxon test was used for g .

Article Snippet: Fibroblasts or co-cultures were trypsinized and directly stained with an equal volume of primary antibody solution (Cell staining buffer, BioLegend) containing eFluor 780 Fixable Viability Dye (Thermo Fisher Scientific; 1:1,000 dilution), TGFβR2-APC (BioLegend; 1:100 dilution), TGFβRIII (Proteintech, 20000-1-AP; 1:100 dilution) or CD31-FITC (BioLegend; 1:100 dilution) for 30 min at 4 C. Cells were washed with cell staining buffer (BioLegend), fixed in 4% paraformaldehyde for 10 min at 20 °C, washed, and stained with AF555 anti-rabbit (Thermo Fisher Scientific) for 30 min at 4 °C for detection of TGFβIII.

Techniques: Enzyme-linked Immunosorbent Assay, Co-Culture Assay, Flow Cytometry, Fluorescence, Expressing, Comparison, Two Tailed Test

Journal: Journal of Translational Medicine

Article Title: SULF1 expression is increased and promotes fibrosis through the TGF-β1/SMAD pathway in idiopathic pulmonary fibrosis

doi: 10.1186/s12967-024-05698-3

Figure Lengend Snippet: A The concentration of TGF-β1 in the serum-free culture supernatants was determined by ELISA. Statistical analysis was conducted using Student’s t-tests (*P < 0.05, **P < 0.01, ***P < 0.001). B The Co-IP with anti-TGFβRIII was performed and the precipitates were analyzed by western blotting with anti-SULF1 or anti-TGF-β1. IgG Co-IP was used as a negative control

Article Snippet: Co-IP was performed using anti-TGFβRIII antibody (#5544, CST) and Protein A/G magnetic beads (B23201, Bimake) according to the manufacturer's protocol.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Co-Immunoprecipitation Assay, Western Blot, Negative Control

Journal: Journal of Translational Medicine

Article Title: SULF1 expression is increased and promotes fibrosis through the TGF-β1/SMAD pathway in idiopathic pulmonary fibrosis

doi: 10.1186/s12967-024-05698-3

Figure Lengend Snippet: Schema depicting proposed pro-fibrotic mechanisms of SULF1 in pulmonary fibrosis. SULF1 catalyzes the release of TGF-β1 bound to TGFβRIII, thereby activating the TGF-β1/SMAD pathway to promote fibrosis. Additionally, TGF-β1 induces SULF1 expression through the TGF-β1/SMAD pathway. There may be a potential positive feedback loop between SULF1 and the TGF-β1/SMAD pathway

Article Snippet: Co-IP was performed using anti-TGFβRIII antibody (#5544, CST) and Protein A/G magnetic beads (B23201, Bimake) according to the manufacturer's protocol.

Techniques: Expressing