gdf10  (MedChemExpress)

Journal: Advanced Science

Article Title: Autocrine GDF10 Inhibits Hepatic Stellate Cell Activation via BMPR2/ALK3 Receptor to Prevent Liver Fibrosis

doi: 10.1002/advs.202500616

Figure Lengend Snippet: Hepatic GDF10 is specifically expressed by HSCs and is downregulated in activated HSCs. A) UMAP visualization of the cell type expresses Gdf10 in the liver ( GSE218299 ). B) Cell type enrichment of GDF10 in human liver (Human Protein Atlas database). C) qPCR analysis of Gdf10 mRNA levels in LSECs, MACs, HSCs, and HCs from the normal, CCl4‐induced fibrotic, and AMLN diet‐induced fibrotic liver. D) IF staining analysis of GDF10 and Desmin protein levels in the liver, scale bars, 20 µm. E,F) UMAP visualization of the Gdf10 and Acta2 mRNA levels in the qHSCs and aHSCs, data from GSE218299 (E) and GSE171904 (F). G) Heat map representation of the indicated genes in HSCs from CCl4 treated or AMLN diet‐fed mice ( GSE134512 ). H,I) qPCR (H) ( n = 6) and IF staining (I) analysis of indicated genes in HSCs isolated from normal and CCl4‐induced fibrotic liver, scale bars, 5 µm. J,K) qPCR (J) ( n = 6) and IF staining (K) analysis of indicated genes in HSCs isolated from normal and AMLN diet‐induced fibrotic liver, scale bars, 5 µm. L,M) qPCR analysis of Desmin ( Des ) mRNA levels (normalized to 36B4) and Gdf10 and Acta2 mRNA levels (normalized to Des ) in fibrotic liver. N) Analysis of DES mRNA levels and GDF10 and ACTA2 mRNA levels (normalized to DES ) in human health and fibrotic liver tissues from dataset GSE159676 . Data are mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 by the two‐tailed Student's t ‐test.

Article Snippet: GDF10‐Fc was purchased from Sino Biological (Cat: 50165‐M01H).

Techniques: Staining, Isolation, Two Tailed Test

Journal: Advanced Science

Article Title: Autocrine GDF10 Inhibits Hepatic Stellate Cell Activation via BMPR2/ALK3 Receptor to Prevent Liver Fibrosis

doi: 10.1002/advs.202500616

Figure Lengend Snippet: GDF10 inhibits TGF‐β‐induced HSC activation. A) qPCR analysis of GDF10 mRNA levels in primary mouse HSCs and LX‐2 cells treated with TGF‐β1 (2 ng mL −1 ), PDGF (20 ng mL −1 ), or vehicle (Veh) for 24 h ( n = 6). B) IF staining analysis of GDF10 protein levels in primary mouse HSCs and LX‐2 cells treated with TGF‐β1 (2 ng mL −1 ) or vehicle for 24 h ( n = 6), the relative fluorescence intensity (RFI) of ACTA2 and GDF10 is also shown, scale bars, 20 µm. C) Heat map representation of the indicated genes in primary human HSCs ( GSE119606 ) and LX‐2 cells ( GSE151251 ) treated with TGF‐β1 or vehicle. D, E) qPCR analysis of Gdf10 and Tgfb1 mRNA levels (D) ( n = 6) and IF staining analysis of GDF10 and ACTA2 protein levels (E) ( n = 3) in HSCs during the culture activation. F) Heat map representation of the indicated genes in HSCs during the culture activation ( GSE116987 ). G–I) qPCR (G) ( n = 6), Western blot (H), and IF staining (I) ( n = 3) analysis of indicated genes in primary mouse HSCs infected with LV‐sh‐Luc or LV‐sh‐Gdf10 for 24 h and then treated with TGF‐β1 (2 ng mL −1 ) or vehicle for another 24 h, scale bars, 20 µm. J–L) qPCR (J) ( n = 6), Western blot (K), and IF staining (L) ( n = 3) analysis of indicated genes in primary mouse HSCs infected with LV‐Con or LV‐Gdf10 for 24 h and then treated with TGF‐β1 (2 ng mL −1 ) or vehicle for another 24 h, scale bars, 20 µm. Data are mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 by the two‐tailed Student's t ‐test (B,D,E), one‐way ANOVA (A, G (left), J (left), I, and L), or two‐way ANOVA (G (right) and J (right)).

Article Snippet: GDF10‐Fc was purchased from Sino Biological (Cat: 50165‐M01H).

Techniques: Activation Assay, Staining, Fluorescence, Western Blot, Infection, Two Tailed Test

Journal: Advanced Science

Article Title: Autocrine GDF10 Inhibits Hepatic Stellate Cell Activation via BMPR2/ALK3 Receptor to Prevent Liver Fibrosis

doi: 10.1002/advs.202500616

Figure Lengend Snippet: Gdf10 knockout promotes HSC activation and exacerbates CCl4‐induced liver fibrosis. A) qPCR analysis of Gdf10 mRNA levels in the HSCs isolated from 12‐week‐old male WT and Gdf10KO mice. B) ELISA analysis of serum GDF10 content in WT and Gdf10KO mice ( n = 6). C) Schematic drawing of the experimental procedure. D) Representative images of H&E, Sirius Red, and ACTA2 IHC staining, scale bars, 50 µm ( n = 3). E) Hydroxyproline assay analysis of total liver collagen content ( n = 6). F‐H) qPCR (F) ( n = 6), Western blot (G), and IF staining (H) ( n = 3) analysis of indicated genes in the liver, scale bars, 50 µm. I, J) Measurement of serum ALT (I) and AST (J) levels of WT and Gdf10KO mice. K, L) qPCR (K) ( n = 3) and IF staining (L) analysis of indicated genes in the primary mouse HSCs, scale bars, 5 µm. Data are mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 by the two‐tailed Student's t ‐test.

Article Snippet: GDF10‐Fc was purchased from Sino Biological (Cat: 50165‐M01H).

Techniques: Knock-Out, Activation Assay, Isolation, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Hydroxyproline Assay, Western Blot, Staining, Two Tailed Test

Journal: Advanced Science

Article Title: Autocrine GDF10 Inhibits Hepatic Stellate Cell Activation via BMPR2/ALK3 Receptor to Prevent Liver Fibrosis

doi: 10.1002/advs.202500616

Figure Lengend Snippet: HSC‐specific Gdf10 transgene inhibits HSC activation and attenuates liver fibrosis in mice. A) qPCR analysis of Gdf10 mRNA levels in the HSCs isolated of 12‐week‐old male Rosa26‐Gdf10 flox/flox (LoxP) and Rosa26‐Gdf10 flox/flox ; Lrat‐Cre (Gdf10TG) mice ( n = 6). B) ELISA analysis of serum GDF10 content in LoxP and Gdf10TG mice ( n = 6). C) Schematic drawing of the experimental procedure. D) Representative images of H&E, Sirius Red, and ACTA2 IHC staining, scale bars, 50 µm ( n = 3). E) Hydroxyproline assay analysis of total liver collagen content ( n = 6). F–H) qPCR (F) ( n = 6), Western blot (G), and IF staining (H) ( n = 3) analysis of indicated genes in the liver, scale bars, 50 µm. I,J) Measurement of serum ALT (I) and AST (J) levels of LoxP and Gdf10TG mice. K,L) qPCR (K) ( n = 3) and IF staining (L) analysis of indicated genes in the primary mouse HSCs, scale bars, 5 µm. Data are mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 by the two‐tailed Student's t ‐test.

Article Snippet: GDF10‐Fc was purchased from Sino Biological (Cat: 50165‐M01H).

Techniques: Activation Assay, Isolation, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Hydroxyproline Assay, Western Blot, Staining, Two Tailed Test

Journal: Advanced Science

Article Title: Autocrine GDF10 Inhibits Hepatic Stellate Cell Activation via BMPR2/ALK3 Receptor to Prevent Liver Fibrosis

doi: 10.1002/advs.202500616

Figure Lengend Snippet: Characterization of the interaction between GDF10 and BMPR2/ALK3. A) Percentage of expression + binding + transfectants of HEK293 cells expressing a single TGF‐β superfamily receptor incubated with GDF10‐Fc. B) Percentage of expression + binding + transfectants of HEK293 cells expressing BMPR2 and type I TGF‐β receptor incubated with GDF10‐Fc. C) HEK293 cells were transfected to express BMPR2 and ALK3 and then incubated with GDF10‐Fc, scale bars, 50 µm. D) Association of BMPR2 with ALK3. HEK293 cells were transfected to express BMPR2‐FLAG and ALK3‐HIS. E) The overall structure of mature GDF10 with the extracellular domains (ECD) of mouse BMPR2 and ALK3 (GDF10: blue‐purple, BMPR2: pink‐green, ALK3: pink). F) Western blot analysis of cell lysates from HEK293 cells transfected to express C‐terminal FLAG‐tagged BMPR2 and FLAG‐tagged ALK3 incubated with GDF10‐Fc and then immunoprecipitated with anti‐FLAG antibody. G) Biacore sensorgrams and binding kinetics were determined by SPR spectroscopy for GDF10 with the ECD of mouse BMPR2 and ALK3. H) qPCR analysis of the expression Bmpr2 and Alk3 in mouse LSECs, MACs, HSCs, and HCs ( n = 6). I) Cell type enrichment of BMPR2 and ALK3 in human liver (Human Protein Atlas database).

Article Snippet: GDF10‐Fc was purchased from Sino Biological (Cat: 50165‐M01H).

Techniques: Expressing, Binding Assay, Incubation, Transfection, Western Blot, Immunoprecipitation, Spectroscopy

Journal: Advanced Science

Article Title: Autocrine GDF10 Inhibits Hepatic Stellate Cell Activation via BMPR2/ALK3 Receptor to Prevent Liver Fibrosis

doi: 10.1002/advs.202500616

Figure Lengend Snippet: GDF10 inhibits HSC activation via the BMPR2/ALK3‐SMAD1/5/8‐SMAD7 signaling pathway. A,B) qPCR (A) ( n = 6) and Western blot (B) analysis of indicated genes in the primary mouse HSCs infected with LV‐sh‐Luc or LV‐sh‐Bmpr2 and LV‐sh‐Alk3 for 24 h and then treated with TGF‐β1 plus Fc or GDF10‐Fc for another 24 h. C) IF staining analysis of ACTA2 expression in HSCs treated as in (A), scale bars, 20 µm ( n = 3). D, E) qPCR ( n = 6) (D) and Western blot (E) analysis of indicated genes in the HSCs treated with TGF‐β1 plus GDF10‐Fc and/or LDN for 24 h. F) IF analysis of ACTA2 stating in HSCs treated as in (D). G) Heat map shows the indicated genes in JS1 cells treated with TGF‐β1 plus LDN and/or GDF10‐Fc for 24 h. H) qPCR (top) ( n = 6) and Western blot (bottom) analysis of SMAD7 mRNA and protein levels in HSCs treated as in (G). I) qPCR (top) ( n = 6) and Western blot (bottom) analysis of SMAD7 mRNA and protein levels in HSCs treated with TGF‐β1 plus DM and/or GDF10‐Fc for 24 h. J) Alignment of the SMAD7 promoter regions containing the GC‐BRE sites (GGCGCC) in the indicated species. K) Luciferase reporter gene assay in JS1 cells transfected with the indicated plasmids and then treated with Fc or GDF10‐Fc. L) ChIP assay showing the recruitment of phosphorylated SMAD1/5/8 to the Smad7 gene promoter in JS1 cells ( n = 3). M,N) qPCR (M) ( n = 6) and Western blot (N) analysis of indicated genes in the primary mouse HSCs infected with LV‐sh‐Luc or LV‐sh‐Smad7 for 24 h, then treated with TGF‐β1 plus Fc or GDF10‐Fc for another 24 h. O) IF staining analysis of ACTA2 expression in HSCs treated as in (M), scale bars, 20 µm ( n = 3). Data are mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 by the two‐tailed Student's t ‐test (K,L), one‐way ANOVA (C,F,H,I,O), or two‐way ANOVA (A,D,M).

Article Snippet: GDF10‐Fc was purchased from Sino Biological (Cat: 50165‐M01H).

Techniques: Activation Assay, Western Blot, Infection, Staining, Expressing, Luciferase, Reporter Gene Assay, Transfection, Two Tailed Test

Journal: Advanced Science

Article Title: Autocrine GDF10 Inhibits Hepatic Stellate Cell Activation via BMPR2/ALK3 Receptor to Prevent Liver Fibrosis

doi: 10.1002/advs.202500616

Figure Lengend Snippet: BMPR2:ALK3‐Fc prevents the antifibrotic effect of GDF10 in the liver. A) Schematic drawing of the experimental procedure in 3‐month‐old male LoxP and Gdf10TG mice. B) Representative images of H&E, Sirius Red, and ACTA2 IHC staining, scale bars, 50 µm ( n = 3). C) Hydroxyproline assay analysis of the total liver collagen of mice treated as in (A) ( n = 6). D,E) qPCR (D) ( n = 6) and Western blot (E) analysis of indicated genes in the liver of mice treated as in (A). F,G) qPCR (F) ( n = 6) and Western blot (G) analysis of indicated genes in the HSCs from mice treated as in (A) ( n = 3). H,I) IF staining (H) and Western blot (I) analysis of indicated protein in the HSCs from mice treated as in (A), scale bars, 5 µm. Data are mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 by the one‐way ANOVA (C,E,G,I) or two‐way ANOVA (D,F).

Article Snippet: GDF10‐Fc was purchased from Sino Biological (Cat: 50165‐M01H).

Techniques: Immunohistochemistry, Hydroxyproline Assay, Western Blot, Staining

Journal: Advanced Science

Article Title: Autocrine GDF10 Inhibits Hepatic Stellate Cell Activation via BMPR2/ALK3 Receptor to Prevent Liver Fibrosis

doi: 10.1002/advs.202500616

Figure Lengend Snippet: Therapeutic administration of GDF10‐Fc ameliorates hepatic fibrosis. A) Schematic drawing of the experimental procedure. B) Representative images of H&E and Sirius Red staining of liver from mice treated as in (A), scale bars, 50 µm ( n = 3). C,D) Measurement of the total liver collagen content by hydroxyproline assay (C) and serum ALT and AST levels (D) in the mice treated as in (A) ( n = 6). E, F) qPCR (E) ( n = 6) and Western blot (F) analysis of indicated genes in the liver of mice treated as in (A). G) Schematic drawing of the experimental procedure and representative images of H&E and Sirius Red staining of liver from AMLN diet‐induced fibrosis, scale bars, 50 µm ( n = 3). H,I) Measurement of the total liver collagen content by hydroxyproline assay (H) and serum ALT and AST levels (I) of the mice treated as in (G) ( n = 6). J,K) qPCR (J) ( n = 6) and Western blot (K) analysis of indicated genes in the liver of mice treated as in (G). L) Schematic drawing of the experimental procedure and representative images of H&E and Sirius Red staining of liver from the MCD diet‐induced fibrosis, scale bars, 50 µm ( n = 3). M,N) Measurement of the total liver collagen content by hydroxyproline assay (M) and serum ALT and AST levels (N) of the mice treated as in (L) ( n = 6). O, P) Western blot (O) and qPCR (P) ( n = 6) analysis of indicated genes in the liver of mice treated as in (L). Q) The proposed model of GDF10 prevents liver fibrosis. Data are mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 by the two‐tailed Student's t ‐test (G–P), one‐way ANOVA (B,C,D,F), or two‐way ANOVA (E).

Article Snippet: GDF10‐Fc was purchased from Sino Biological (Cat: 50165‐M01H).

Techniques: Staining, Hydroxyproline Assay, Western Blot, Two Tailed Test