Journal: Cell Death Discovery

Article Title: Interplay between ALK2 R206H mutant receptor and autophagy signaling regulates receptor stability and its chondrogenic functions

doi: 10.1038/s41420-025-02393-0

Figure Lengend Snippet: A Flow cytometry analysis in ATDC5 cells overexpressing ALK2 WT or mutant ALK2 R206H receptors in regular medium (5% FCS) and serum starvation (starvation) conditions. GFP-positive and RFP-positive cells were detected by fluorescence emitted under 488 nm and 561 nm excitation lasers, respectively. Representative histograms of a single experiment with median fluorescence intensity of GFP and RFP are shown. Cumulative plots of six independent experiments show mean ± SD of GFP/RFP fluorescence ratio in serum starvation conditions versus regular medium (DMEM F12 5% FBS). Statistical analysis: One-way ANOVA (Tukey’s multiple comparison) test was performed (ns = not significant; ** P ≤ 0.01; **** P ≤ 0.0001) ( B ) Representative immunoblotting and relative densitometric analysis ( n = 3) on total protein extracts derived from ATDC5 cells grown in regular medium (5% FBS) or in serum starvation condition (starvation). Tubulin and HSP90 were used as loading control. Statistical analysis: unpaired t -test (ns = not significant; ** P ≤ 0.01; **** P ≤ 0.0001) ( C ) Confocal quantification of the number of autophagic vesicles in ATDC5 cells in starvation condition after loading with 50 µM MDC. Data analysis of MDC spot number was performed using the Harmony software (ver 4.5) of the Opera Phenix high-content system. Scale bar, 10 μm. Statistical analysis: unpaired t- test (**** P ≤ 0.0001).

Article Snippet: For immunoblotting, 20–30 μg of protein extract were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), blotted on nitrocellulose membrane, and stained with the specific primary antibodies: anti-Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E,Cell Signaling Technology), anti-p62 (SQSTM1) (PM045; MBL International), anti-GAPDH (D16H11) (Cell Signaling Technology), anti-LC3B (Sigma–Aldrich, L7543), anti-NanoLuc (Promega Corporation, N700A), anti-DDK (FLAG) (Origene, TA50011-100), anti ALK2/ACVR1 (SinoBiological.

Techniques: Flow Cytometry, Mutagenesis, Fluorescence, Comparison, Western Blot, Derivative Assay, Control, Software

Journal: Cell Death Discovery

Article Title: Interplay between ALK2 R206H mutant receptor and autophagy signaling regulates receptor stability and its chondrogenic functions

doi: 10.1038/s41420-025-02393-0

Figure Lengend Snippet: A Histograms show the mRNA expression for the indicated genes expressed as fold increase (mean ± SD) from three independent experiments in ATDC5 cells overexpressing ALK2 WT or mutant ALK2 R206H in normoxic (21% O 2 ) and hypoxic conditions (1% O 2 ,16 h). GAPDH mRNA was used to normalize data. B Immunoblotting and corresponding densitometric analysis with the indicated antibodies of ATDC5 cells overexpressing ALK2 WT or mutant ALK2 R206H in normoxic (21% O 2 ) and hypoxic conditions (1% O 2 ,16 h) ( C ) Histograms show the mRNA expression for the indicated gene expressed as fold increase (mean ± SD) from three independent experiments in ATDC5 cells overexpressing ALK2 WT or mutant ALK2 R206H in normoxic (21% O 2 ) and hypoxic conditions (1% O 2 ,16 h). GAPDH mRNA was used to normalize data. D , E Immunoblotting and relative densitometric analysis with the indicated antibodies of U2OS cells overexpressing ALK2 WT or mutant ALK2 R206H DDK tagged, in normoxic (21% O 2 ) and hypoxic conditions (1% O 2 ,16 h) and treated, as indicated, with 20 μM CQ (2 h). F Confocal microscopy analysis using anti-DDK (red) in U2OS cells expressing ALK2 WT or mutant ALK2 R206H DDK tagged receptors treated or not, as indicated, with chloroquine (CQ, 20 μM, 2 h) in normoxic (21% O 2 ) and hypoxic conditions (1% O 2 ,16 h). DNA was counterstained with DAPI (blue). Scale bar 10 µm. DDK fluorescence intensity was measured by using Fiji software (ImageJ). At least 50 cells from 3 independent experiments were analyzed unpaired t -test (Welch’s correction) was performed. Data are presented as mean ± SD of at least three independent experiments. Symbols (dots) represent individual cells. Statistical analysis: unpaired (ns = not significant; * P ≤ 0.05; **P ≤ 0.01; *** P < 0.001 ****P ≤ 0.0001).

Article Snippet: For immunoblotting, 20–30 μg of protein extract were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), blotted on nitrocellulose membrane, and stained with the specific primary antibodies: anti-Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E,Cell Signaling Technology), anti-p62 (SQSTM1) (PM045; MBL International), anti-GAPDH (D16H11) (Cell Signaling Technology), anti-LC3B (Sigma–Aldrich, L7543), anti-NanoLuc (Promega Corporation, N700A), anti-DDK (FLAG) (Origene, TA50011-100), anti ALK2/ACVR1 (SinoBiological.

Techniques: Expressing, Mutagenesis, Western Blot, Confocal Microscopy, Fluorescence, Software

Journal: Cell Death Discovery

Article Title: Interplay between ALK2 R206H mutant receptor and autophagy signaling regulates receptor stability and its chondrogenic functions

doi: 10.1038/s41420-025-02393-0

Figure Lengend Snippet: A Representative images and relative quantification of EGFP-LC3-labeled autophagosomes colocalization with ALK2-DDK receptor. U2OS EGFP-LC3 cells expressing ALK2 WT or mutant ALK2 R206H DDK tagged receptors were treated with chloroquine (CQ, 20 μM, 2 h) in hypoxic conditions (1% O 2 ,16 h). Images were acquired by spinning disk confocal microscopy and analyzed for colocalization of EGFP-LC3 (green) and DDK (red) dots using the ComeDet plugin of the Fiji ImageJ software. Data are presented as mean ± SD and normalized on total EGFP-LC3 dots. DNA was counterstained with DAPI (blue). Insets show a 3-fold enlargement of the boxed areas. Scale bar, 10 μm is shown. At least 50 cells from 3 independent experiments were analyzed unpaired t -test (Welch’s correction) was performed. (ns = not significant; * P ≤ 0.05; ** P ≤ 0.01; *** P < 0.001 **** P ≤ 0.0001). B Immunoblotting and relative densitometric analysis, numbers report the densitometric values of band intensity, with the indicated antibodies of U2OS cells overexpressing ALK2 WT or mutant ALK2 R206H in normoxic (21% O 2 ) and hypoxic conditions (1% O 2 ,16 h) and stably infected with lentiviral construct expressing shATG5 RNA interference or expressing an empty vector as control (shCTRL).

Article Snippet: For immunoblotting, 20–30 μg of protein extract were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), blotted on nitrocellulose membrane, and stained with the specific primary antibodies: anti-Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E,Cell Signaling Technology), anti-p62 (SQSTM1) (PM045; MBL International), anti-GAPDH (D16H11) (Cell Signaling Technology), anti-LC3B (Sigma–Aldrich, L7543), anti-NanoLuc (Promega Corporation, N700A), anti-DDK (FLAG) (Origene, TA50011-100), anti ALK2/ACVR1 (SinoBiological.

Techniques: Labeling, Expressing, Mutagenesis, Confocal Microscopy, Software, Western Blot, Stable Transfection, Infection, Construct, Plasmid Preparation, Control

Journal: Cell Death Discovery

Article Title: Interplay between ALK2 R206H mutant receptor and autophagy signaling regulates receptor stability and its chondrogenic functions

doi: 10.1038/s41420-025-02393-0

Figure Lengend Snippet: A Immunoblotting and relative densitometric analysis with the indicated antibodies of ALK2 R206H -DDK U2OS cells treated or not, at different time points, with activin A (100 ng/ml) and Rapamycin (Rapa, 100 ng/ml) in serum starvation conditions. Histograms show the mean ± SD of at least three independent experiments. Statistical analysis: unpaired t -test (ns = not significant; * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001 **** P ≤ 0.0001). B Representative fluorescence images of U2OS cells expressing mutant ALK2 R206H DDK tagged receptors treated or not, as indicated, with Rapamycin (Rapa, 100 ng/ml, 5 h) and/or chloroquine (CQ, 20 μM, 2 h), immunostained with anti-LAMP2 (red) and anti-DDK (green) antibodies and counterstained with DNA (DAPI, blue). Scale bar 10 µm. Images were acquired by spinning disk confocal microscopy and analyzed for DDK fluorescence intensity using the Fiji ImageJ software. Data are presented as median with interquartile range. N ≥ 20 cells per sample from two independent experiments. Symbols represent individual cells. Insets show a 3-fold enlargement of the boxed areas. Statistical analysis: one-way ANOVA (Tukey’s multiple comparison) test was performed (** P ≤ 0.01; **** P ≤ 0.0001). C Flow cytometry analysis of ALK2 R206H ATDC5 cells cultured in serum starvation conditions, left untreated (NT) or treated with activin A (100 ng /ml) alone (DMSO), or in combination with Chloroquine (CQ, 20 µM, 2 h), Rapamycin (Rapa, 100 ng/ml, 24 h) for 24 h. Representative histograms show the median fluorescence intensity (MFI) of fluorescence GFP and RFP emitted as a result of excitation with blue (488 nm) and yellow (561 nm) lasers, respectively. Cumulative plots report the mean ± SD of GFP/RFP fluorescence ratio versus non-treated (NT) cells ( n = 3). Statistical analysis: one-way ANOVA (Tukey’s multiple comparison) test was performed (ns = not significant; ** P ≤ 0.01).

Article Snippet: For immunoblotting, 20–30 μg of protein extract were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), blotted on nitrocellulose membrane, and stained with the specific primary antibodies: anti-Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E,Cell Signaling Technology), anti-p62 (SQSTM1) (PM045; MBL International), anti-GAPDH (D16H11) (Cell Signaling Technology), anti-LC3B (Sigma–Aldrich, L7543), anti-NanoLuc (Promega Corporation, N700A), anti-DDK (FLAG) (Origene, TA50011-100), anti ALK2/ACVR1 (SinoBiological.

Techniques: Western Blot, Fluorescence, Expressing, Mutagenesis, Confocal Microscopy, Software, Comparison, Flow Cytometry, Cell Culture

Journal: Cell Death Discovery

Article Title: Interplay between ALK2 R206H mutant receptor and autophagy signaling regulates receptor stability and its chondrogenic functions

doi: 10.1038/s41420-025-02393-0

Figure Lengend Snippet: A Chondrogenic differentiation micromass assay in ATDC5 ALK2 R206H cells and in ATDC5 ALK2 WT cells. Cells were incubated for 21 days in differentiation medium containing or not (NT), activin A (100 ng/ml) and treated, as indicated, with autophagy inhibitors Chloroquine (CQ, 20 μM), 3-Methyladenine (3-MA, 20 μM) or autophagy inducers Rapamycin (Rapa, 100 ng/ml), Spermidine (SPD, 20 μM) or DMSO. Representative images of Alcian blue staining. Histograms representing Alcian blue quantification measured by absorbance at 595 nm after solubilization with guanidine hydrochloride. All results represented the mean ± SD of at least three independent experiments. Statistical analysis: One-way ANOVA (Tukey’s multiple comparison) test was performed (** P ≤ 0.01; *** P < 0.001 **** P ≤ 0.0001). B Real-time PCR of Col10a1 mRNA in ATDC5 cells treated or not with activin A for 21 days in micromass cultures. mRplp0 was used to normalize data. All results represented the mean ± SD of at least three independent experiments. Statistical analysis: unpaired t- test (** P ≤ 0.01; *** P < 0.001 **** P ≤ 0.0001). C Histogram showing measurement of ALP activity in ATDC5 cells treated or not as indicated for 5 days in micromass culture. 5 × 10 4 cells were homogenized in 1 ml of assay Buffer, diluted 1:10 in assay Buffer, and 80 μl was used to measure ALP activity. Assays were performed following the fluorimetric kit protocol (MAK411, Sigma–Aldrich) and measured at O.D. 405 nm reading with Varioskan LUX Plate Reader. All results represented the mean ± SD of at least three independent experiments. Statistical analysis: A one-way ANOVA (Tukey’s multiple comparison) test was performed (** P ≤ 0.01; *** P < 0.001). Chondrogenic differentiation micromass assay in ATDC5 ALK2 R206H cells stable expressing shRNA for ATG4 (shATG4) ( D ) or ATDC5 ALK2 WT and ALK2 R206H cells stable expressing shRNA for Rubicon (shRubicon) ( E ), upon lentiviral infection. A lentiviral empty vector was used as control (shCTRL). Cells were incubated for 21 days in differentiation medium containing or not (NT), activin A (100 ng/ml) and treated as indicated with Rapamycin (Rapa 100 ng/ml) or DMSO. Representative images of Alcian blue staining and relative histograms showing Alcian blue quantification measured by absorbance at 595 nm after solubilization with guanidine hydrochloride. All results represented the mean ± SD of at least three independent experiments for ( D ) and from six independent experiments for ( E ). Statistical analysis: One-way ANOVA (Tukey’s multiple comparison) test was performed (* P ≤ 0.05; ** P ≤ 0.01, **** P ≤ 0.0001).

Article Snippet: For immunoblotting, 20–30 μg of protein extract were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), blotted on nitrocellulose membrane, and stained with the specific primary antibodies: anti-Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E,Cell Signaling Technology), anti-p62 (SQSTM1) (PM045; MBL International), anti-GAPDH (D16H11) (Cell Signaling Technology), anti-LC3B (Sigma–Aldrich, L7543), anti-NanoLuc (Promega Corporation, N700A), anti-DDK (FLAG) (Origene, TA50011-100), anti ALK2/ACVR1 (SinoBiological.

Techniques: Incubation, Staining, Comparison, Real-time Polymerase Chain Reaction, Activity Assay, Expressing, shRNA, Infection, Plasmid Preparation, Control

Journal: Cell Death Discovery

Article Title: Interplay between ALK2 R206H mutant receptor and autophagy signaling regulates receptor stability and its chondrogenic functions

doi: 10.1038/s41420-025-02393-0

Figure Lengend Snippet: Schematic illustration showing a simplified model of the interplay between the mutant ALK2 R206H receptor (without showing the type I/type II receptors tetramer) and the mTOR/autophagy signaling in the regulation of the balance between receptor degradation and chondrogenesis upon hypoxic conditions or activin A stimulation.

Article Snippet: For immunoblotting, 20–30 μg of protein extract were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), blotted on nitrocellulose membrane, and stained with the specific primary antibodies: anti-Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E,Cell Signaling Technology), anti-p62 (SQSTM1) (PM045; MBL International), anti-GAPDH (D16H11) (Cell Signaling Technology), anti-LC3B (Sigma–Aldrich, L7543), anti-NanoLuc (Promega Corporation, N700A), anti-DDK (FLAG) (Origene, TA50011-100), anti ALK2/ACVR1 (SinoBiological.

Techniques: Mutagenesis

Journal: The EMBO Journal

Article Title: GNAS/PKA signaling promotes aberrant osteochondral differentiation of Gli1 + tendon sheath progenitors

doi: 10.1038/s44318-025-00553-7

Figure Lengend Snippet: ( A ) Schematic diagram depicting the generation of Acvr1 R206H/+ flox mice using CRISPR-Cas9 technology. ( B ) microCT showing spontaneous HO formation around the joints of Prrx1-Cre ; Acvr1 R206H/+ mice, but not Prrx1-Cre mice. ( C , D ) Representative microCT image ( C ) and statistical analysis ( D ) of HO formation in the tibial muscle of Tek -Cre ; Acvr1 R206H/+ mice, but not Tek -Cre mice after CTX injury ( n = 4 per group). **** P = 6.05e-7. ( E , F ) Representative HE staining ( E) and safranine O staining ( F ) images of the injured tendon of Gli1-CreER T2 and Gli1-CreER T2 ; Acvr1 R206H/+ mice at 7 dpi. Scale bar, 200 μm. ( G ) Statistical analysis of Safranine O + region in injured tendon of Gli1-CreER T2 and Gli1-CreER T2 ; Acvr1 R206H/+ mice at 7 dpi ( n = 5 per group). **** P = 2.28e-6. ( H ) Statistical analysis of HO volume in the injured tendon of Gli1-CreER T2 and Gli1-CreER T2 ; Acvr1 R206H/+ mice at 56 dpi ( n = 5 per group). **** P = 1.29e-5. ( I ) Representative microCT images of HO in the injured tendon of Gli1-CreER T2 and Gli1-CreER T2 ; Acvr1 R206H/+ mice at 56 dpi. ( J , K ) Statistical analysis ( J ) and representative immunofluorescence images ( K ) of Gli1 + cells in the injured tendon of Gli1-CreER T2 ; Ai9 and Gli1-CreER T2 ; Ai9; Acvr1 R206H/+ mice at 3, 5, and 7 dpi ( n = 5 per group). * P = 3.86e-2, ** P = 3.82e-3, **** P = 2.73e-4. Scale bar, 100 μm. Data are represented as the mean ± SD. All P values were determined by unpaired Student’s t test.

Article Snippet: Anti-mouse ACVR1 , Proteintech , Cat#67417-1-Ig.

Techniques: CRISPR, Staining, Immunofluorescence

Journal: The EMBO Journal

Article Title: GNAS/PKA signaling promotes aberrant osteochondral differentiation of Gli1 + tendon sheath progenitors

doi: 10.1038/s44318-025-00553-7

Figure Lengend Snippet: ( A , B ) Representative immunofluorescence and statistical analysis of GLI1 and ACVR1 in injured site of Gli1-CreER T2 ; Ai9 and Gli1-CreER T2 ; Ai9; Acvr1 R206H/+ mice at 5 dpi ( n = 5 per group). P = 5.18e-1. N.S. indicated no significance. Scale bar, 100:μm. ( C , D ) Representative immunofluorescence and statistical analysis of GLI1 and p-SMAD1/5 in injured site of Gli1-CreER T2 ; Ai9 and Gli1-CreER T2 ; Ai9; Acvr1 R206H/+ mice at 5 dpi ( n = 5 per group). **** P = 6.89e-6. Scale bar, 100 μm. ( E ) Sanger sequencing showed that the STOP cassette preceding the Acvr1 R206H/+ mutation was disrupted. Data is presented as mean ± SD. All P values were determined by unpaired Student’s t test.

Article Snippet: Anti-mouse ACVR1 , Proteintech , Cat#67417-1-Ig.

Techniques: Immunofluorescence, Sequencing, Mutagenesis

Journal: The EMBO Journal

Article Title: GNAS/PKA signaling promotes aberrant osteochondral differentiation of Gli1 + tendon sheath progenitors

doi: 10.1038/s44318-025-00553-7

Figure Lengend Snippet: ( A , B ) Representative immunofluorescence images ( A ) and statistical analysis of frequency ( B ) of GLI1 - / FMOD - , GLI1 + / FMOD - , GLI1 + / FMOD + , GLI1 - / FMOD + cells in the injured site of FOP model mice at 3 dpi ( n = 5 per group). P values from left to right: ** P = 2.27e-3, **** P = 1.17e-12, *** P = 4.59e-4, **** P = 6.94e-13, **** P = 2.49e-11. Scale bar, 200 or 20 μm. ( C, D ) Representative immunofluorescence images ( C ) and statistical analysis of frequency ( D ) of GLI1 - / SOX9 - , GLI1 + /SOX9 − , GLI1 − /SOX9 + , GLI1 + / SOX9 + cells in the injured site of FOP model mice at 5 dpi ( n = 5 per group). P values from left to right: * P = 2.52e-2, * P = 1.85e-2, **** P = 3.43e-5, *** P = 4.18e-4. Scale bar, 200 or 20 μm. ( E , F ) Representative immunofluorescence images ( E ) and statistical analysis of frequency ( F ) of GLI1 − /RUNX2 - , GLI1 + /RUNX2 − , GLI1 − /RUNX2 + , GLI1 + /RUNX2 + cells in the injured site of FOP model mice at 5 dpi ( n = 5 per group). P values from left to right: **** P = 5.56e-5, **** P = 1.32e-6, ** P = 1.12e-3, **** P = 1.73e-5. Scale bar, 200 or 20 μm. ( G , H ) Representative immunofluorescence images ( G ) and statistical analysis of frequency ( H ) of GLI1 - /COL2 - , GLI1 + / COL2 − , GLI1 + / COL2 + , GLI1 - / COL2 + cells in the injured site of FOP model mice at 7 dpi ( n = 5 per group). P values from left to right: **** P = 1.52e-10, **** P = 2.90e-11, **** P = 6.99e-11. Scale bar, 200 or 20 μm. ( I ) Representative immunofluorescence images showing Gli1 + tendon sheath progenitors differentiate into osteocytes at 14 dpi in FOP model mice. Scale bar, 200 or 20 μm. ( J ) Statistical analysis of the frequency of GLI1 - /OCN - , GLI1 + /OCN + , GLI1 - /OCN + , GLI1 + /OCN - cells in the injured site of FOP model mice at 14 dpi ( n = 5 per group). P values from left to right: **** P = 8.87e-11, **** P = 9.49e-7, **** P = 4.35e-10, **** P = 3.85e-6, **** P = 6.35e-5. ( K , L ) Representative Safranine O staining images ( K ) and statistical analysis ( L ) of chondrocyte region in the injured tendon of Gli1 - CreER T2 ; Acvr1 R206H/+ and Gli1 - CreER T2 ; Acvr1 R206H/+ ; DTA mice ( n = 5 per group). *** P = 3.27e-4. Scale bar, 200 μm. ( M , N ) Representative microCT images ( M ) and statistical analysis ( N ) of HO in the injured tendon of Gli1 - Cre ERT2 ; Acvr1 R206H/+ and Gli1 - Cre ERT2 ; Acvr1 R206H/+ ; DTA mice at 63 dpi ( n = 5 per group). **** P = 7.86e-7. Data are presented as mean ± SD. All P values were determined by One-way ANOVA with Bonferroni post hoc test ( B , D , F , H , J ) and unpaired Student’s t test ( L , N ).

Article Snippet: Anti-mouse ACVR1 , Proteintech , Cat#67417-1-Ig.

Techniques: Immunofluorescence, Staining

Journal: The EMBO Journal

Article Title: GNAS/PKA signaling promotes aberrant osteochondral differentiation of Gli1 + tendon sheath progenitors

doi: 10.1038/s44318-025-00553-7

Figure Lengend Snippet: ( A – D ) Representative immunofluorescence images of GNAS expression in uninjured tendons ( A ) and injured sites of Gli1 - CreER T2 ; Ai9 mice at 5 ( B ), 7 ( C ) and 14 ( D ) dpi. Scale bar, 50 μm. ( E , F ) Statistical analysis of GNAS + region ( E ) and GNAS + /GLI1 + region ( F ) in uninjured tendons and injured sites of Gli1 - CreER T2 ; Acvr1 R206H/+ mice at 5, 7 and 14 dpi ( n = 5 per group). P values from left to right: **** P = 8.29e-10, **** P = 1.16e-8, **** P = 1.57e-8. **** P = 2.36e-9, **** P = 1.71e-6, **** P = 9.06e-6. ( G , H ) Statistical analysis of p-PKA substrate + region ( G ) and p-PKA substrate + /GLI1 + region ( H ) in uninjured tendons and injured sites of Gli1 - CreER T2 ; Acvr1 R206H/+ mice at 5, 7 and 14 dpi ( n = 5 per group). P values from left to right: **** P = 1.42e-6, **** P = 7.23e-6, **** P = 1.53e-5. **** P = 1.96e-5, ** P = 2.63e-3, **** P = 6.44e-4. ( I – L ) Representative immunofluorescence images of p-PKA substrate expression in uninjured tendons ( I ) and injured sites of Gli1 - CreER T2 ; Acvr1 R206H/+ mice at 5 ( J ), 7 ( K ) and 14 ( L ) dpi. Scale bar, 50 μm. Data is presented as mean ± SD. All P values were determined by unpaired Student’s t test.

Article Snippet: Anti-mouse ACVR1 , Proteintech , Cat#67417-1-Ig.

Techniques: Immunofluorescence, Expressing

Journal: The EMBO Journal

Article Title: GNAS/PKA signaling promotes aberrant osteochondral differentiation of Gli1 + tendon sheath progenitors

doi: 10.1038/s44318-025-00553-7

Figure Lengend Snippet: ( A , B ) Representative immunofluorescence images ( A ) and statistical analysis ( B ) of SOX9 + region in injured site from tenotomized mice following vehicle and NF449 treatment ( n = 5 per group). *** P = 8.94e-4. Scale bar, 10 μm. ( C , D ) Representative immunofluorescence images ( C ) and statistical analysis ( D ) of RUNX2 + region in injured site from tenotomized mice following vehicle and NF449 treatment ( n = 5 per group). * P = 1.37e-2. Scale bar, 10 μm. ( E , F ) Representative Safranine O staining images ( E ) and statistical analysis ( F ) of Safranine O + region in injured site from tenotomized mice following vehicle and NF449 treatment ( n = 5 per group). ** P = 1.85e-3. Scale bar, 100 μm. ( G , H ) Representative microCT images ( G ) and statistical analysis ( H ) of HO volume in injured site from tenotomized mice following vehicle and NF449 treatment ( n = 5 per group). *** P = 1.39e-4. ( I , J ) Representative Safranine O staining images ( I ) and statistical analysis ( J ) of Safranine O + region in injured site from tenotomized Gli1-CreER T2 and Gli1-CreER T2 ; Prkaca f/f mice ( n = 5 per group). *** P = 1.18e-4. Scale bar, 100 μm. ( K , L ) Representative microCT images ( K ) and statistical analysis ( L ) of HO volume in injured site from tenotomized Gli1-CreER T2 and Gli1-CreER T2 ; Prkaca f/f mice ( n = 5 per group). *** P = 5.11e-4. ( M , N ) Representative Safranine O staining images ( M ) and statistical analysis ( N ) of Safranine O + region in injured site from tenotomized Gli1-CreER T2 ; Acvr1 R206H/+ mice following vehicle and NF449 treatment ( n = 5 per group). **** P = 1.5e-7. Scale bar, 100 μm. ( O , P ) Representative microCT images ( O ) and statistical analysis ( P ) of HO volume in injured site from tenotomized Gli1-CreER T2 ; Acvr1 R206H/+ mice following vehicle and NF449 treatment ( n = 5 per group). **** P = 3.87e-6. ( Q , R ) Representative Safranine O staining images ( Q ) and statistical analysis ( R ) of Safranine O + region in injured site from tenotomized Gli1-CreER T2 ; Acvr1 R206H/+ and Gli1-CreER T2 ; Acvr1 R206H/+ ; Prkaca f/f mice at 7 dpi ( n = 5 per group). **** P = 6.2e-7. Scale bar, 100 μm. ( S , T ) Representative microCT images ( S ) and statistical analysis ( T ) of HO volume in injured site from tenotomized Gli1-CreER T2 ; Acvr1 R206H/+ and Gli1-CreER T2 ; Acvr1 R206H/+ ; Prkaca f/f mice at 14 dpi ( n = 5 per group). *** P = 1.79e-4. Data are presented as mean ± SD. All P values were determined by unpaired Student’s t test.

Article Snippet: Anti-mouse ACVR1 , Proteintech , Cat#67417-1-Ig.

Techniques: Immunofluorescence, Staining

Journal: European Journal of Histochemistry : EJH

Article Title: Expression of bone morphogenetic protein signaling pathway players in the jejunum and colon of adult rats

doi: 10.4081/ejh.2025.4174

Figure Lengend Snippet: Immunohistochemical localization of ACVR1 ( a,b,c ) and BMPR2 ( d,e,f ) receptors in rat jejunum. e ) The arrows show strong immunostaining in the lamina propria; the asterisk designates the longitudinal muscularis layer. To determine the localization of BMP receptors, we performed immunohistochemical staining on jejunum sections from 6 different rats. Sections were examined by light microscopy (Leica DM 2000) using 20x and 40x objectives. Representative images are shown. Scale bars: 50 μm.

Article Snippet: ACVR1 , 1:1000 , ProteinTech - 67417-1 - Ig.

Techniques: Immunohistochemical staining, Immunostaining, Staining, Light Microscopy

Journal: European Journal of Histochemistry : EJH

Article Title: Expression of bone morphogenetic protein signaling pathway players in the jejunum and colon of adult rats

doi: 10.4081/ejh.2025.4174

Figure Lengend Snippet: Immunohistochemical localization of ACVR1 ( a,b,c ) and BMPR2 ( d,e,f ) receptors in rat colon. b ) The asterisk shows surface colonocytes. To determine the localization of BMP receptors, we performed immunohistochemical staining on colon sections from 6 different rats. Sections were examined by light microscopy (Leica DM 2000) using 20x and 40x objectives. Representative images are shown. Scale bars: 50 μm.

Article Snippet: ACVR1 , 1:1000 , ProteinTech - 67417-1 - Ig.

Techniques: Immunohistochemical staining, Staining, Light Microscopy