Journal: NPJ Microgravity

Article Title: BMP9 attenuates microgravity-related disuse osteoporosis by modulating TGFβ and BMP signaling

doi: 10.1038/s41526-025-00510-y

Figure Lengend Snippet: A Alkaline phosphatase staining. B Quantification of alkaline phosphatase enzyme activity. C Osteogenic gene expression following infection with Adv-BMP9 or control virus in the presence of 10 ng/ml TGFβ1 ( n = 3). D Phosphorylation of Smads in marrow stromal cells following infection with Adv-BMP9 or control virus in the presence of 10 ng/ml TGFβ1, as detected by western blot. E BMP-pSmad1/5/8 signaling target gene expression in marrow stromal cells following infection with Adv-BMP9 or control virus in the presence of 10 ng/ml TGFβ1 ( n = 3). ns no significance, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Bone marrow stromal cells were extracted using the previously mentioned method , infected with Adv-BMP9 or control virus, and then treated with osteogenic induction medium (OM) containing TGFβ1 at concentrations of 1, 5, and 10 ng/mL (Topscience, China) to assess its impact on osteogenic function.

Techniques: Staining, Activity Assay, Gene Expression, Infection, Control, Virus, Phospho-proteomics, Western Blot, Targeted Gene Expression

Journal: NPJ Microgravity

Article Title: BMP9 attenuates microgravity-related disuse osteoporosis by modulating TGFβ and BMP signaling

doi: 10.1038/s41526-025-00510-y

Figure Lengend Snippet: A Phosphorylation of Smads in primary osteocytes following infection with Adv-BMP9 or control virus in the presence of 10 ng/ml TGFβ1, as detected by western blot. B BMP-pSmad1/5/8 signaling target gene expression in primary osteocytes following infection with Adv-BMP9 or control virus in the presence of 10 ng/ml TGFβ1 ( n = 3). C Intracellular pH changes in primary osteocytes following infection with Adv-BMP9 or control virus in the presence of 10 ng/ml TGFβ1, normalized to control group. D Expression of osteoclastic genes in primary osteocytes following infection with Adv-BMP9 or control virus in the presence of 10 ng/ml TGFβ1 ( n = 3). ns no significance, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Bone marrow stromal cells were extracted using the previously mentioned method , infected with Adv-BMP9 or control virus, and then treated with osteogenic induction medium (OM) containing TGFβ1 at concentrations of 1, 5, and 10 ng/mL (Topscience, China) to assess its impact on osteogenic function.

Techniques: Phospho-proteomics, Infection, Control, Virus, Western Blot, Targeted Gene Expression, Expressing

Journal: Comprehensive Physiology

Article Title: Disrupting BMP / TGF ‐β Signaling: Modulation of AQP1 and TGFB1 in Human Pulmonary Microvascular Endothelial Cells

doi: 10.1002/cph4.70066

Figure Lengend Snippet: Effects of silencing the AQP1 gene and exogenous administration of BMP9 on AQP1 and BMP/TGF‐β signaling molecules in human pulmonary microvascular endothelial cells. HPMECs were silenced for the AQP1 gene and the relative mRNA and protein expression of AQP1 (A, B), BMPR2 (C, D), TGFBR1 (E, F), and TGFB1 (G, H) were estimated before and after the exogenous administration of BMP9. Relative mRNA expression is shown in dot plots (dots, individual values; line in the middle, median values; lower and upper lines, 25th and 75th percentiles) (A, n = 6; C, n = 7; E, n = 7; G, n = 6). Protein expression was analyzed by SDS‐PAGE and immunoblotting, and relative expression was estimated by densitometry using β‐tubulin as a loading control. Relative protein expression is shown with bar plots (mean ± SEM; dots, individual values) (B, n = 4; D, n = 4; F, n = 4; H, n = 4). Representative expression of AQP1 (B), BMPR2 (D), TGFBR1 (F), and TGFB1 (H) (upper panels) and β‐tubulin (lower panels) proteins in the AQP1 ‐silenced HPMEC homogenates. Specificity (siRNA negative control), efficiency ( AQP1 siRNA), and the effect of BMP9 exogenous administration on the non‐transfected controls were tested each time to ensure the consistency and reproducibility across the independent experiments. Statistical analysis was performed using the Mann–Whitney test. *, p < 0.05; **, p < 0.01 compared to the non‐transfected control HPMECs.

Article Snippet: HPMECs were treated with BMP9 (5 ng/mL) (OriGene, Rockville, MD, USA) 24 h post‐transfection.

Techniques: Expressing, SDS Page, Western Blot, Control, Negative Control, Transfection, MANN-WHITNEY

Journal: Comprehensive Physiology

Article Title: Disrupting BMP / TGF ‐β Signaling: Modulation of AQP1 and TGFB1 in Human Pulmonary Microvascular Endothelial Cells

doi: 10.1002/cph4.70066

Figure Lengend Snippet: Effects of silencing the TGFB1 gene and exogenous administration of BMP9 on BMP/TGF‐β signaling molecules and AQP1 in human pulmonary microvascular endothelial cells. HPMECs were silenced for the TGFB1 gene and the relative mRNA and protein expression of TGFB1 (A, B), TGFBR1 (C, D), AQP1 (E, F), and BMPR2 (G, H) were estimated before and after the exogenous administration of BMP9. Relative mRNA expression is shown in dot plots (dots, individual values; line in the middle, median values; lower and upper lines, 25th and 75th percentiles) (A, n = 5; C, n = 6; E, n = 4; G, n = 7). Protein expression was analyzed by SDS‐PAGE and immunoblotting, and relative expression was estimated by densitometry using β‐tubulin or Actin as a loading control. Relative protein expression is shown with bar plots (mean ± SEM; dots, individual values) (B, n = 4; D, n = 7; F, n = 4; H, n = 6). Representative expression of TGFB1 (B), TGFBR1 (D), AQP1 (F), and BMPR2 (H) (upper panels) and β‐tubulin (lower panels) proteins in the TGFB1 ‐silenced HPMEC homogenates. Specificity (siRNA negative control), efficiency ( TGFB1 siRNA), and the effect of BMP9 exogenous administration on the non‐transfected controls were tested each time to ensure consistency and reproducibility across the independent experiments. Statistical analysis was performed using the Mann–Whitney test. *, p < 0.05; **, p < 0.01 compared to the non‐transfected control HPMECs.

Article Snippet: HPMECs were treated with BMP9 (5 ng/mL) (OriGene, Rockville, MD, USA) 24 h post‐transfection.

Techniques: Expressing, SDS Page, Western Blot, Control, Negative Control, Transfection, MANN-WHITNEY

Journal: Comprehensive Physiology

Article Title: Disrupting BMP / TGF ‐β Signaling: Modulation of AQP1 and TGFB1 in Human Pulmonary Microvascular Endothelial Cells

doi: 10.1002/cph4.70066

Figure Lengend Snippet: Effects of silencing the TGFB1 gene and exogenous administration of BMP9 on BMP/TGF‐β ligands in human pulmonary microvascular endothelial cells. HPMECs were silenced for the TGFB1 gene, and the relative mRNA and protein expression of BMP9 (A, B) and BMP10 (C, D) were estimated. BMP10 expression was estimated before and after the exogenous administration of BMP9. Relative mRNA expression is shown in dot plots (dots, individual values; line in the middle, median values; lower and upper lines, 25th and 75th percentiles) (A, n = 6; C, n = 5). Protein expression was analyzed by SDS‐PAGE and immunoblotting, and relative expression was estimated by densitometry using β‐tubulin or Actin as a loading control. Relative protein expression is shown with bar plots (mean ± SEM; dots, individual values) (B, n = 4; D, n = 5). Representative expression of BMP9 (B), and BMP10 (D) (upper panels) and β‐tubulin/Actin (lower panels) proteins in the TGFB1 ‐silenced HPMEC homogenates. Specificity (siRNA negative control), efficiency ( TGFB1 siRNA), and the effect of BMP9 exogenous administration on the non‐transfected controls were tested each time to ensure consistency and reproducibility across the independent experiments. Statistical analysis was performed using the Mann–Whitney test. *, p < 0.05 compared to the non‐transfected control HPMECs.

Article Snippet: HPMECs were treated with BMP9 (5 ng/mL) (OriGene, Rockville, MD, USA) 24 h post‐transfection.

Techniques: Expressing, SDS Page, Western Blot, Control, Negative Control, Transfection, MANN-WHITNEY

Journal: Communications Biology

Article Title: LRIG proteins regulate lipid metabolism via BMP signaling and affect the risk of type 2 diabetes

doi: 10.1038/s42003-020-01613-w

Figure Lengend Snippet: a Wild-type (WT) and Lrig -null (TKO) MEFs expressing the BMP reporter plasmid pGL3-BRE-luciferase were treated with the indicated concentrations of BMP4 for three hours. Thereafter, the cells were lysed, and the luciferase activity was analyzed and normalized to the control. b – d Wild-type and Lrig -null MEFs were stimulated with various concentrations of BMP4 ( b ), BMP6 ( c ), or BMP9 ( d ) for one hour followed by immunocytofluorescence analysis of nuclear phospho-Smad1/5 (pSmad1/5). e , f Wild-type and Lrig1 -null MEFs ( e ) or wild-type and Lrig3 -null MEFs ( f ) were stimulated with various concentrations of BMP4 for one hour followed by nuclear pSmad1/5 analysis. g – i Western blot analyses of canonical BMP4 signaling through pSmad1/5 and noncanonical BMP signaling through phosphorylated p38 (pp38). Wild-type and Lrig -null MEFs were stimulated with the indicated concentrations of BMP4 for one hour followed by cell lysis and Western blot analysis. Uncropped blots are shown in Supplementary Fig. . g Representative Western blots showing pSmad1/5, pp38, total Smad1, total p38, and the loading control actin. h Quantification of the pSmad1/5/actin ratios. i Quantification of the pp38/actin ratios. j Gene expression levels were analyzed in wild-type (WT) and Lrig -null (TKO) MEFs via RNA sequencing (RNAseq). The apparent number of RNAseq reads for respective gene is indicated. All the values in a – f , h and i represent the means of four biological replicates that were analyzed by three experimental repeats each. j The values represent the means of four biological replicates that were analyzed once. Error bars represent the standard deviations of means from four biological replicates. ns P > 0.05, * P < 0.05, ** P < 0.01, *** P < 0.001 (Student’s t -test).

Article Snippet: Cells were starved in serum-free cell culture medium for one hour and then stimulated with BMP4, recombinant human BMP6 (PeproTech Nordic, catalog # 120-06), recombinant human GDF2/BMP9 (PeproTech Nordic, catalog # 120-07), or TGF-β1 for one hour.

Techniques: Expressing, Plasmid Preparation, Luciferase, Activity Assay, Western Blot, Lysis, RNA Sequencing Assay