Journal: Redox Biology

Article Title: SIRT3-mediated deacetylation of FoxM1 prevents pulmonary fibrosis via modulating the activation of pulmonary fibroblasts

doi: 10.1016/j.redox.2026.104108

Figure Lengend Snippet: FoxM1 is highly expressed in fibroblasts of fibrotic lung tissues. (A, B) qPCR (n = 9) and Western blot (n = 6) analysis of the expression of FoxM1 in normal and IPF lung tissues. ∗ P < 0.05. (C) qPCR analysis of the mRNA levels of FoxM1 in the lung tissues from BLM-treated mice. n = 3, ∗ P < 0.05. (D) Western blot analysis of the protein levels of FoxM1, CTHRC1, α-SMA, and Collagen I in the lung tissues from bleomycin (BLM)-treated mice. n = 3, ∗ P < 0.05. (E) The Pearson's correlation analysis of COL1A1 expression with FoxM1 expression based on the RNA-seq results of GSE24206 from GEO database. (F) The Pearson's correlation analysis of Ashcroft score with FoxM1 expression in the lung tissues from BLM-treated mice. (G) Representative images of co-immunostaining for α-SMA and FoxM1 in IPF lung tissues. White arrows indicate double-positive cells. (H) Representative images of co-immunostaining for α-SMA and FoxM1 in the lung tissues from BLM-treated mice. White arrows indicate double-positive cells. (I) Western blot analysis of FoxM1 expression in pulmonary fibroblasts isolated from mice subjected to BLM treatment. n = 3, ∗ P < 0.05. (J) Western blot analysis of FoxM1, CTHRC1, α-SMA, and Collagen I expression in pulmonary fibroblasts treated with TGF-β1. n = 3, ∗ P < 0.05. All data were presented as the means ± SEM. Paired t -test (A, B, I) and one-way ANOVA with Tukey's post-hoc test (C, D, J) were used for statistical analysis.

Article Snippet: To explore the role of FoxM1 in pulmonary fibrosis, mice were treated with RCM-1 (MedChem Express, San Diego, CA, USA), which is an inhibitor of FoxM1 [ ].

Techniques: Western Blot, Expressing, RNA Sequencing, Immunostaining, Isolation

Journal: Redox Biology

Article Title: SIRT3-mediated deacetylation of FoxM1 prevents pulmonary fibrosis via modulating the activation of pulmonary fibroblasts

doi: 10.1016/j.redox.2026.104108

Figure Lengend Snippet: Enhanced FoxM1 expression is responsible for the resistance of fibroblasts to FasL-mediated apoptosis . (A) Caspase 3 activity analysis in pulmonary fibroblasts isolated from bleomycin (BLM)-treated mice following FasL treatment. (B) Cell viability assessment using the CCK-8 assay in pulmonary fibroblasts isolated from BLM-treated mice after FasL treatment. (C) Caspase 3 activity analysis in TGF-β1-treated pulmonary fibroblasts, along with or without FasL treatment. (D) Cell viability assessment using the CCK-8 assay in TGF-β1-treated pulmonary fibroblasts, along with or without FasL treatment. (E) Western blot analysis of FoxM1 expression in pulmonary fibroblasts transfected with or without LV-FoxM1. (F) Western blot analysis of FoxM1 expression in pulmonary fibroblasts transfected with or without FoxM1 siRNA (si-FoxM1). (G, H) Caspase 3 activity and cell viability assessment in pulmonary fibroblasts transfected with or without LV-FoxM1, following with or without FasL treatment. (I, J) Caspase 3 activity and cell viability assessment in pulmonary fibroblasts transfected with or without si-FoxM1, following with or without FasL treatment. All data (n = 3, ∗ P < 0.05) were presented as the means ± SEM. Paired t -test was used for statistical analysis.

Article Snippet: To explore the role of FoxM1 in pulmonary fibrosis, mice were treated with RCM-1 (MedChem Express, San Diego, CA, USA), which is an inhibitor of FoxM1 [ ].

Techniques: Expressing, Activity Assay, Isolation, CCK-8 Assay, Western Blot, Transfection

Journal: Redox Biology

Article Title: SIRT3-mediated deacetylation of FoxM1 prevents pulmonary fibrosis via modulating the activation of pulmonary fibroblasts

doi: 10.1016/j.redox.2026.104108

Figure Lengend Snippet: Impairing nuclear translocation of FoxM1 suppresses fibroblast activation and protects mice from bleomycin-induced pulmonary fibrosis. (A) Western blot analysis was performed to assess the nuclear expression levels of FoxM1 in pulmonary fibroblasts isolated from BLM-treated mice. n = 3, ∗ P < 0.05. (B) Western blot analysis of nuclear FoxM1, CTHRC1, α-SMA, and Collagen I expression in TGF-β1-treated pulmonary fibroblasts accompany with or without RCM-1 treatment. n = 3, ∗ P < 0.05. (C, D) EdU assay for the proliferation of TGF-β1-treated pulmonary fibroblasts accompany with or without RCM-1 treatment. n = 3, ∗ P < 0.05. (E) Hematoxylin–eosin (H&E) and masson's trichrome staining for the lung tissues from BLM-treated mice injected with or without RCM-1. (F, G) The ashcroft score (n = 6, ∗ P < 0.05.) and hydroxyproline contents (n = 6, ∗ P < 0.05.) in the lung tissues of BLM-treated mice injected with or without RCM-1. (H) The survival of BLM-treated mice injected with or without RCM-1. n = 18. (I) Western blot analysis of FoxM1, CTHRC1, α-SMA, and Collagen I expression in the lung tissues from BLM-treated mice injected with or without RCM-1. n = 3, ∗ P < 0.05. All data were presented as the means ± SEM. Paired t -test (A) and one-way ANOVA with Tukey's post-hoc test (B, D, F–I) were used for statistical analysis.

Article Snippet: To explore the role of FoxM1 in pulmonary fibrosis, mice were treated with RCM-1 (MedChem Express, San Diego, CA, USA), which is an inhibitor of FoxM1 [ ].

Techniques: Translocation Assay, Activation Assay, Western Blot, Expressing, Isolation, EdU Assay, Staining, Injection

Journal: Redox Biology

Article Title: SIRT3-mediated deacetylation of FoxM1 prevents pulmonary fibrosis via modulating the activation of pulmonary fibroblasts

doi: 10.1016/j.redox.2026.104108

Figure Lengend Snippet: Acetylation of FoxM1 is required for the activation of pulmonary fibroblasts. (A, B) Western blot analysis of FoxM1 expression in the cytoplasm and nucleus of CHX-treated pulmonary fibroblasts along with or without MG132 treatment at indicated time. n = 3, ∗ P < 0.05. (C) Western blot analysis of FoxM1 expression in CHX-treated pulmonary fibroblasts along with or without TGF-β1 treatment. n = 3, ∗ P < 0.05. (D) Western blot analysis of the acetylation levels of FoxM1 in pulmonary fibroblasts isolated from bleomycin (BLM)-treated mice. n = 3, ∗ P < 0.05. (E) Western blot analysis of the acetylation levels of FoxM1 in pulmonary fibroblasts treated with or without TGF-β1. n = 3, ∗ P < 0.05. (F, G) Western blot analysis of FoxM1 acetylation, CTHRC1, α-SMA, and Collagen I expression in pulmonary fibroblasts treated with TSA (50 nM), or NAM (1 mM) for 24 h, or not. n = 3, ∗ P < 0.05. All data were presented as the means ± SEM. Paired t -test (D, E) and one-way ANOVA with Tukey's post-hoc test (B, C, G) were used for statistical analysis.

Article Snippet: To explore the role of FoxM1 in pulmonary fibrosis, mice were treated with RCM-1 (MedChem Express, San Diego, CA, USA), which is an inhibitor of FoxM1 [ ].

Techniques: Activation Assay, Western Blot, Expressing, Isolation

Journal: Redox Biology

Article Title: SIRT3-mediated deacetylation of FoxM1 prevents pulmonary fibrosis via modulating the activation of pulmonary fibroblasts

doi: 10.1016/j.redox.2026.104108

Figure Lengend Snippet: Sirt3-dependent deacetylation of FoxM1 regulates the stability of FoxM1. (A) The Pearson's correlation analysis of COL1A1 expression with SIRTs expression based on the RNA-seq results of GSE2052 from GEO database. (B) Western blot analysis of SIRT3 expression in the lung tissues from bleomycin (BLM)-treated mice. n = 3, ∗ P < 0.05. (C) Western blot analysis of the acetylation levels of FoxM1 in SIRT3 flox/flox mice intratracheally injected with AAV-Cre. n = 3, ∗ P < 0.05. (D) Western blot analysis was performed to assess the acetylation status of FoxM1 in pulmonary fibroblasts following transfection with Sirt3 siRNA (si-Sirt3). n = 3, ∗ P < 0.05. (E) Western blot analysis of FoxM1 expression in CHX-treated pulmonary fibroblasts transfected with or without si-Sirt3. n = 3, ∗ P < 0.05. (F) Western blot analysis of CTHRC1, α-SMA and Collagen I expression in pulmonary fibroblasts transfected with or without si-Sirt3. n = 3, ∗ P < 0.05. (G) EdU assay for the proliferation of pulmonary fibroblasts transfected with or without si-Sirt3. n = 3, ∗ P < 0.05. (H) Western blot analysis of FoxM1 acetylation, CTHRC1, α-SMA, and Collagen I expression in TGF-β1-treated pulmonary fibroblasts transfected with or without LV-Sirt3. n = 3, ∗ P < 0.05. All data were presented as the means ± SEM. Paired t -test (B-D, F, G) and one-way ANOVA with Tukey's post-hoc test (E, H) were used for statistical analysis.

Article Snippet: To explore the role of FoxM1 in pulmonary fibrosis, mice were treated with RCM-1 (MedChem Express, San Diego, CA, USA), which is an inhibitor of FoxM1 [ ].

Techniques: Expressing, RNA Sequencing, Western Blot, Injection, Transfection, EdU Assay

Journal: Redox Biology

Article Title: SIRT3-mediated deacetylation of FoxM1 prevents pulmonary fibrosis via modulating the activation of pulmonary fibroblasts

doi: 10.1016/j.redox.2026.104108

Figure Lengend Snippet: Sirt3 knockdown accelerates BLM-induced pulmonary fibrosis via activation pulmonary fibroblasts in vivo. (A) Hematoxylin–eosin (H&E) and masson's trichrome staining for the lung tissues from SIRT3 flox/flox mice or BLM-treated SIRT3 flox/flox mice that intratracheally injected with or without AAV-Cre. (B, C) The ashcroft score (n = 6, ∗ P < 0.05.) and the hydroxyproline contents (n = 6, ∗ P < 0.05.) in the lung tissues of mice treated as in A. (D) Western blot analysis of CTHRC1, SIRT3, FoxM1, α-SMA and Collagen I expression in the lung tissues from mice treated as in A n = 3, ∗ P < 0.05. (E) Western blot analysis of CTHRC1, SIRT3, FoxM1, α-SMA and Collagen I expression in pulmonary fibroblasts isolated from mice treated as in A n = 3, ∗ P < 0.05. (F, G) EdU assay for the proliferation of pulmonary fibroblasts isolated from mice treated as in A. All data were presented as the means ± SEM. One-way ANOVA with Tukey's post-hoc test was used for statistical analysis.

Article Snippet: To explore the role of FoxM1 in pulmonary fibrosis, mice were treated with RCM-1 (MedChem Express, San Diego, CA, USA), which is an inhibitor of FoxM1 [ ].

Techniques: Knockdown, Activation Assay, In Vivo, Staining, Injection, Western Blot, Expressing, Isolation, EdU Assay

Journal: Redox Biology

Article Title: SIRT3-mediated deacetylation of FoxM1 prevents pulmonary fibrosis via modulating the activation of pulmonary fibroblasts

doi: 10.1016/j.redox.2026.104108

Figure Lengend Snippet: Nicotinamide riboside protects mice from bleomycin-induced pulmonary fibrosis via activation of SIRT3. (A) Western blot analysis of CTHRC1, SIRT3, FoxM1, α-SMA and Collagen I expression in TGF-β1-treated pulmonary fibroblasts accompany with or without NR treatment. n = 3, ∗ P < 0.05. (B) Cell viability assessment using the CCK-8 assay in pulmonary fibroblasts treated as in A n = 6, ∗ P < 0.05. (C) The survival of BLM-treated mice oral gavaged with or without NR. n = 18. (D) Hematoxylin–eosin (H&E) and masson's trichrome staining for the lung tissues from mice treated as in C. (E) The ashcroft score of mice treated as in C n = 6, ∗ P < 0.05. (F) Western blot analysis of CTHRC1, SIRT3, FoxM1, α-SMA and Collagen I expression in the lung tissues from mice treated as in C n = 3, ∗ P < 0.05. (G) Western blot analysis of CTHRC1, SIRT3, FoxM1, α-SMA and Collagen I expression in pulmonary fibroblasts isolated from the lung tissues of mice treated as in C n = 3, ∗ P < 0.05. All data were presented as the means ± SEM. One-way ANOVA with Tukey's post-hoc test was used for statistical analysis.

Article Snippet: To explore the role of FoxM1 in pulmonary fibrosis, mice were treated with RCM-1 (MedChem Express, San Diego, CA, USA), which is an inhibitor of FoxM1 [ ].

Techniques: Activation Assay, Western Blot, Expressing, CCK-8 Assay, Staining, Isolation

Journal: Redox Biology

Article Title: SIRT3-mediated deacetylation of FoxM1 prevents pulmonary fibrosis via modulating the activation of pulmonary fibroblasts

doi: 10.1016/j.redox.2026.104108

Figure Lengend Snippet: FoxM1 is highly expressed in fibroblasts of fibrotic lung tissues. (A, B) qPCR (n = 9) and Western blot (n = 6) analysis of the expression of FoxM1 in normal and IPF lung tissues. ∗ P < 0.05. (C) qPCR analysis of the mRNA levels of FoxM1 in the lung tissues from BLM-treated mice. n = 3, ∗ P < 0.05. (D) Western blot analysis of the protein levels of FoxM1, CTHRC1, α-SMA, and Collagen I in the lung tissues from bleomycin (BLM)-treated mice. n = 3, ∗ P < 0.05. (E) The Pearson's correlation analysis of COL1A1 expression with FoxM1 expression based on the RNA-seq results of GSE24206 from GEO database. (F) The Pearson's correlation analysis of Ashcroft score with FoxM1 expression in the lung tissues from BLM-treated mice. (G) Representative images of co-immunostaining for α-SMA and FoxM1 in IPF lung tissues. White arrows indicate double-positive cells. (H) Representative images of co-immunostaining for α-SMA and FoxM1 in the lung tissues from BLM-treated mice. White arrows indicate double-positive cells. (I) Western blot analysis of FoxM1 expression in pulmonary fibroblasts isolated from mice subjected to BLM treatment. n = 3, ∗ P < 0.05. (J) Western blot analysis of FoxM1, CTHRC1, α-SMA, and Collagen I expression in pulmonary fibroblasts treated with TGF-β1. n = 3, ∗ P < 0.05. All data were presented as the means ± SEM. Paired t -test (A, B, I) and one-way ANOVA with Tukey's post-hoc test (C, D, J) were used for statistical analysis.

Article Snippet: The primary antibodies used were: FoxM1, α-SMA, SIRT3 and Lamin B1 (Selleckchem, Houston, USA), Collagen I (Proteintech, Wuhan, China) and GAPDH (ABclonal, Wuhan, China).

Techniques: Western Blot, Expressing, RNA Sequencing, Immunostaining, Isolation

Journal: Redox Biology

Article Title: SIRT3-mediated deacetylation of FoxM1 prevents pulmonary fibrosis via modulating the activation of pulmonary fibroblasts

doi: 10.1016/j.redox.2026.104108

Figure Lengend Snippet: Enhanced FoxM1 expression is responsible for the resistance of fibroblasts to FasL-mediated apoptosis . (A) Caspase 3 activity analysis in pulmonary fibroblasts isolated from bleomycin (BLM)-treated mice following FasL treatment. (B) Cell viability assessment using the CCK-8 assay in pulmonary fibroblasts isolated from BLM-treated mice after FasL treatment. (C) Caspase 3 activity analysis in TGF-β1-treated pulmonary fibroblasts, along with or without FasL treatment. (D) Cell viability assessment using the CCK-8 assay in TGF-β1-treated pulmonary fibroblasts, along with or without FasL treatment. (E) Western blot analysis of FoxM1 expression in pulmonary fibroblasts transfected with or without LV-FoxM1. (F) Western blot analysis of FoxM1 expression in pulmonary fibroblasts transfected with or without FoxM1 siRNA (si-FoxM1). (G, H) Caspase 3 activity and cell viability assessment in pulmonary fibroblasts transfected with or without LV-FoxM1, following with or without FasL treatment. (I, J) Caspase 3 activity and cell viability assessment in pulmonary fibroblasts transfected with or without si-FoxM1, following with or without FasL treatment. All data (n = 3, ∗ P < 0.05) were presented as the means ± SEM. Paired t -test was used for statistical analysis.

Article Snippet: The primary antibodies used were: FoxM1, α-SMA, SIRT3 and Lamin B1 (Selleckchem, Houston, USA), Collagen I (Proteintech, Wuhan, China) and GAPDH (ABclonal, Wuhan, China).

Techniques: Expressing, Activity Assay, Isolation, CCK-8 Assay, Western Blot, Transfection

Journal: Redox Biology

Article Title: SIRT3-mediated deacetylation of FoxM1 prevents pulmonary fibrosis via modulating the activation of pulmonary fibroblasts

doi: 10.1016/j.redox.2026.104108

Figure Lengend Snippet: Impairing nuclear translocation of FoxM1 suppresses fibroblast activation and protects mice from bleomycin-induced pulmonary fibrosis. (A) Western blot analysis was performed to assess the nuclear expression levels of FoxM1 in pulmonary fibroblasts isolated from BLM-treated mice. n = 3, ∗ P < 0.05. (B) Western blot analysis of nuclear FoxM1, CTHRC1, α-SMA, and Collagen I expression in TGF-β1-treated pulmonary fibroblasts accompany with or without RCM-1 treatment. n = 3, ∗ P < 0.05. (C, D) EdU assay for the proliferation of TGF-β1-treated pulmonary fibroblasts accompany with or without RCM-1 treatment. n = 3, ∗ P < 0.05. (E) Hematoxylin–eosin (H&E) and masson's trichrome staining for the lung tissues from BLM-treated mice injected with or without RCM-1. (F, G) The ashcroft score (n = 6, ∗ P < 0.05.) and hydroxyproline contents (n = 6, ∗ P < 0.05.) in the lung tissues of BLM-treated mice injected with or without RCM-1. (H) The survival of BLM-treated mice injected with or without RCM-1. n = 18. (I) Western blot analysis of FoxM1, CTHRC1, α-SMA, and Collagen I expression in the lung tissues from BLM-treated mice injected with or without RCM-1. n = 3, ∗ P < 0.05. All data were presented as the means ± SEM. Paired t -test (A) and one-way ANOVA with Tukey's post-hoc test (B, D, F–I) were used for statistical analysis.

Article Snippet: The primary antibodies used were: FoxM1, α-SMA, SIRT3 and Lamin B1 (Selleckchem, Houston, USA), Collagen I (Proteintech, Wuhan, China) and GAPDH (ABclonal, Wuhan, China).

Techniques: Translocation Assay, Activation Assay, Western Blot, Expressing, Isolation, EdU Assay, Staining, Injection

Journal: Redox Biology

Article Title: SIRT3-mediated deacetylation of FoxM1 prevents pulmonary fibrosis via modulating the activation of pulmonary fibroblasts

doi: 10.1016/j.redox.2026.104108

Figure Lengend Snippet: Acetylation of FoxM1 is required for the activation of pulmonary fibroblasts. (A, B) Western blot analysis of FoxM1 expression in the cytoplasm and nucleus of CHX-treated pulmonary fibroblasts along with or without MG132 treatment at indicated time. n = 3, ∗ P < 0.05. (C) Western blot analysis of FoxM1 expression in CHX-treated pulmonary fibroblasts along with or without TGF-β1 treatment. n = 3, ∗ P < 0.05. (D) Western blot analysis of the acetylation levels of FoxM1 in pulmonary fibroblasts isolated from bleomycin (BLM)-treated mice. n = 3, ∗ P < 0.05. (E) Western blot analysis of the acetylation levels of FoxM1 in pulmonary fibroblasts treated with or without TGF-β1. n = 3, ∗ P < 0.05. (F, G) Western blot analysis of FoxM1 acetylation, CTHRC1, α-SMA, and Collagen I expression in pulmonary fibroblasts treated with TSA (50 nM), or NAM (1 mM) for 24 h, or not. n = 3, ∗ P < 0.05. All data were presented as the means ± SEM. Paired t -test (D, E) and one-way ANOVA with Tukey's post-hoc test (B, C, G) were used for statistical analysis.

Article Snippet: The primary antibodies used were: FoxM1, α-SMA, SIRT3 and Lamin B1 (Selleckchem, Houston, USA), Collagen I (Proteintech, Wuhan, China) and GAPDH (ABclonal, Wuhan, China).

Techniques: Activation Assay, Western Blot, Expressing, Isolation

Journal: Redox Biology

Article Title: SIRT3-mediated deacetylation of FoxM1 prevents pulmonary fibrosis via modulating the activation of pulmonary fibroblasts

doi: 10.1016/j.redox.2026.104108

Figure Lengend Snippet: Sirt3-dependent deacetylation of FoxM1 regulates the stability of FoxM1. (A) The Pearson's correlation analysis of COL1A1 expression with SIRTs expression based on the RNA-seq results of GSE2052 from GEO database. (B) Western blot analysis of SIRT3 expression in the lung tissues from bleomycin (BLM)-treated mice. n = 3, ∗ P < 0.05. (C) Western blot analysis of the acetylation levels of FoxM1 in SIRT3 flox/flox mice intratracheally injected with AAV-Cre. n = 3, ∗ P < 0.05. (D) Western blot analysis was performed to assess the acetylation status of FoxM1 in pulmonary fibroblasts following transfection with Sirt3 siRNA (si-Sirt3). n = 3, ∗ P < 0.05. (E) Western blot analysis of FoxM1 expression in CHX-treated pulmonary fibroblasts transfected with or without si-Sirt3. n = 3, ∗ P < 0.05. (F) Western blot analysis of CTHRC1, α-SMA and Collagen I expression in pulmonary fibroblasts transfected with or without si-Sirt3. n = 3, ∗ P < 0.05. (G) EdU assay for the proliferation of pulmonary fibroblasts transfected with or without si-Sirt3. n = 3, ∗ P < 0.05. (H) Western blot analysis of FoxM1 acetylation, CTHRC1, α-SMA, and Collagen I expression in TGF-β1-treated pulmonary fibroblasts transfected with or without LV-Sirt3. n = 3, ∗ P < 0.05. All data were presented as the means ± SEM. Paired t -test (B-D, F, G) and one-way ANOVA with Tukey's post-hoc test (E, H) were used for statistical analysis.

Article Snippet: The primary antibodies used were: FoxM1, α-SMA, SIRT3 and Lamin B1 (Selleckchem, Houston, USA), Collagen I (Proteintech, Wuhan, China) and GAPDH (ABclonal, Wuhan, China).

Techniques: Expressing, RNA Sequencing, Western Blot, Injection, Transfection, EdU Assay

Journal: Redox Biology

Article Title: SIRT3-mediated deacetylation of FoxM1 prevents pulmonary fibrosis via modulating the activation of pulmonary fibroblasts

doi: 10.1016/j.redox.2026.104108

Figure Lengend Snippet: Sirt3 knockdown accelerates BLM-induced pulmonary fibrosis via activation pulmonary fibroblasts in vivo. (A) Hematoxylin–eosin (H&E) and masson's trichrome staining for the lung tissues from SIRT3 flox/flox mice or BLM-treated SIRT3 flox/flox mice that intratracheally injected with or without AAV-Cre. (B, C) The ashcroft score (n = 6, ∗ P < 0.05.) and the hydroxyproline contents (n = 6, ∗ P < 0.05.) in the lung tissues of mice treated as in A. (D) Western blot analysis of CTHRC1, SIRT3, FoxM1, α-SMA and Collagen I expression in the lung tissues from mice treated as in A n = 3, ∗ P < 0.05. (E) Western blot analysis of CTHRC1, SIRT3, FoxM1, α-SMA and Collagen I expression in pulmonary fibroblasts isolated from mice treated as in A n = 3, ∗ P < 0.05. (F, G) EdU assay for the proliferation of pulmonary fibroblasts isolated from mice treated as in A. All data were presented as the means ± SEM. One-way ANOVA with Tukey's post-hoc test was used for statistical analysis.

Article Snippet: The primary antibodies used were: FoxM1, α-SMA, SIRT3 and Lamin B1 (Selleckchem, Houston, USA), Collagen I (Proteintech, Wuhan, China) and GAPDH (ABclonal, Wuhan, China).

Techniques: Knockdown, Activation Assay, In Vivo, Staining, Injection, Western Blot, Expressing, Isolation, EdU Assay

Journal: Redox Biology

Article Title: SIRT3-mediated deacetylation of FoxM1 prevents pulmonary fibrosis via modulating the activation of pulmonary fibroblasts

doi: 10.1016/j.redox.2026.104108

Figure Lengend Snippet: Nicotinamide riboside protects mice from bleomycin-induced pulmonary fibrosis via activation of SIRT3. (A) Western blot analysis of CTHRC1, SIRT3, FoxM1, α-SMA and Collagen I expression in TGF-β1-treated pulmonary fibroblasts accompany with or without NR treatment. n = 3, ∗ P < 0.05. (B) Cell viability assessment using the CCK-8 assay in pulmonary fibroblasts treated as in A n = 6, ∗ P < 0.05. (C) The survival of BLM-treated mice oral gavaged with or without NR. n = 18. (D) Hematoxylin–eosin (H&E) and masson's trichrome staining for the lung tissues from mice treated as in C. (E) The ashcroft score of mice treated as in C n = 6, ∗ P < 0.05. (F) Western blot analysis of CTHRC1, SIRT3, FoxM1, α-SMA and Collagen I expression in the lung tissues from mice treated as in C n = 3, ∗ P < 0.05. (G) Western blot analysis of CTHRC1, SIRT3, FoxM1, α-SMA and Collagen I expression in pulmonary fibroblasts isolated from the lung tissues of mice treated as in C n = 3, ∗ P < 0.05. All data were presented as the means ± SEM. One-way ANOVA with Tukey's post-hoc test was used for statistical analysis.

Article Snippet: The primary antibodies used were: FoxM1, α-SMA, SIRT3 and Lamin B1 (Selleckchem, Houston, USA), Collagen I (Proteintech, Wuhan, China) and GAPDH (ABclonal, Wuhan, China).

Techniques: Activation Assay, Western Blot, Expressing, CCK-8 Assay, Staining, Isolation

Journal: bioRxiv

Article Title: A Wnt-responsive fibrocartilage progenitor system coordinates postnatal mandibular condylar cartilage growth

doi: 10.64898/2026.03.25.714159

Figure Lengend Snippet: (A) UMAP visualization after regression of cell-cycle effects showing that the MC-progenitor cluster remains preserved following cell-cycle correction. (B) Feature plots showing expression of representative cell-cycle–associated genes ( Mki67, Pcna, Top2a, Cdk1, Ccnb1 ) and Foxm1. (C) Violin plots showing S phase and G2/M phase cell-cycle scores across clusters. (D) RNA velocity analysis using scVelo showing directional transcriptional flows from the MC-progenitor cluster toward fibrocartilage and chondrocartilage populations.

Article Snippet: Pharmacological treatments were performed using the Foxm1 inhibitor RCM-1, the β-catenin inhibitor XAV-939, or a canonical Wnt pathway activator (Wnt agonist 1) (all from Selleck Chemicals, Japan).

Techniques: Expressing

Journal: bioRxiv

Article Title: A Wnt-responsive fibrocartilage progenitor system coordinates postnatal mandibular condylar cartilage growth

doi: 10.64898/2026.03.25.714159

Figure Lengend Snippet: (A) Differential gene expression analysis was performed comparing H2B–EGFP –positive and H2B–EGFP –negative cells within the MC-progenitor cluster identified by single-cell RNA sequencing. Genes are ranked by statistical significance. Foxm1 is significantly enriched in the H2B–EGFP –positive population, whereas Tgfb1 is enriched in the H2B–EGFP –negative population, indicating divergent transcriptional programs associated with Wnt activity. (B) Feature plots were generated to visualize expression of representative genes across mandibular condylar cartilage populations. Wnt-responsive cells show enriched expression of Foxm1 and IGF signaling–related genes ( Igf1 , Igf1r , Igfbp4 , Igfbp7 ), whereas Wnt-low populations express Tgfb1 and related factors ( Igf2r , Igfbp5 , Igfbp6 ), supporting distinct signaling states. (C) Western blot analysis was performed in isolated Wnt-responsive cells transfected with control vector or constitutively active β-catenin (S33Y). Cells were stimulated with recombinant IGF1 for the indicated time points. β-catenin activation enhances Foxm1 expression and downstream mitogenic signaling, including ERK and IGF1R phosphorylation, indicating that β-catenin promotes proliferative signaling responses. (D) Co-immunoprecipitation was performed to assess interaction between β-catenin and Foxm1. Cell lysates immunoprecipitated with anti–β-catenin antibody show enrichment of Foxm1 compared with control IgG, indicating a physical association between β-catenin and Foxm1. (E,F) Histological and immunofluorescence analyses were performed on mandibular condyles from control and Axin2 CreERT2 ;Ctnnb1 fl/+ ;Foxm1 fl/+ compound heterozygous mice at P42. H&E staining reveals reduced fibrocartilage thickness, and Ki67 staining shows decreased proliferative activity, indicating cooperative effects of β-catenin and Foxm1 in maintaining fibrocartilage proliferation. Scale bar, 100 μm. (G) Quantification of fibrocartilage thickness and Ki67-positive cells was performed. Compound heterozygous mice show reduced fibrocartilage thickness and decreased proliferation compared with controls. Data are presented as mean ± s.d. Each dot represents one biologically independent animal. (H,I) Histological and immunofluorescence analyses were performed on mandibular condyles from control and Axin2 CreERT2 ;Foxm1 fl/fl mice at P42. Foxm1 deletion results in marked condylar hypoplasia and reduced proliferative activity, indicating a critical role for Foxm1 in fibrocartilage growth. Scale bar, 100 μm. (J) Quantification of cartilage thickness and proliferative indices was performed in Foxm1 conditional knockout mice. Foxm1 deficiency significantly reduces cartilage growth and proliferation. Data are presented as mean ± s.d. Each dot represents one biologically independent animal. Statistical significance was assessed using two-tailed Student’s t-test. n.s., not significant; **P < 0.01; ****P < 0.0001. Abbreviations: sz, superficial zone; fc, fibrocartilage zone; cc, chondrocartilage zone.

Article Snippet: Pharmacological treatments were performed using the Foxm1 inhibitor RCM-1, the β-catenin inhibitor XAV-939, or a canonical Wnt pathway activator (Wnt agonist 1) (all from Selleck Chemicals, Japan).

Techniques: Gene Expression, Single Cell, RNA Sequencing, Activity Assay, Generated, Expressing, Western Blot, Isolation, Transfection, Control, Plasmid Preparation, Recombinant, Activation Assay, Phospho-proteomics, Immunoprecipitation, Immunofluorescence, Staining, Knock-Out, Two Tailed Test

Journal: bioRxiv

Article Title: A Wnt-responsive fibrocartilage progenitor system coordinates postnatal mandibular condylar cartilage growth

doi: 10.64898/2026.03.25.714159

Figure Lengend Snippet: (A) RNAscope in situ hybridization showing Foxm1 transcript localization within the fibrocartilage compartment of the mandibular condyle. (B) RNAscope detection of Igf1 transcripts enriched in the superficial region of the fibrocartilage layer. (C) Violin plot comparing Foxm1 expression between H2B-EGFP –positive and H2B-EGFP –negative cells within the MC-progenitor cluster. Scale bars: 100 μm.

Article Snippet: Pharmacological treatments were performed using the Foxm1 inhibitor RCM-1, the β-catenin inhibitor XAV-939, or a canonical Wnt pathway activator (Wnt agonist 1) (all from Selleck Chemicals, Japan).

Techniques: RNAscope, In Situ Hybridization, Expressing

Journal: bioRxiv

Article Title: A Wnt-responsive fibrocartilage progenitor system coordinates postnatal mandibular condylar cartilage growth

doi: 10.64898/2026.03.25.714159

Figure Lengend Snippet: Isolated Wnt-responsive cells were cultured in vitro and treated with the Wnt pathway inhibitor XAV-939 (1 μM or 5 μM), the Foxm1 inhibitor RCM-1 (4 μM or 20 μM), or the Wnt agonist Wnt agonist 1 (0.2 nM or 1 nM). Cell proliferation was quantified by measuring total cell number after treatment. Pharmacological inhibition of canonical Wnt signaling using XAV-939 significantly reduced cell expansion compared with control cultures. Similarly, inhibition of Foxm1 using RCM-1 markedly suppressed proliferation. In contrast, activation of canonical Wnt signaling by Wnt agonist 1 increased cell numbers relative to controls. These findings support a cooperative role for canonical Wnt signaling and Foxm1 in promoting proliferative capacity of Wnt-responsive cells in vitro . Bars represent mean ± s.d. Each dot represents one biologically independent sample. Statistical significance was determined by one-way ANOVA followed by multiple comparisons testing. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

Article Snippet: Pharmacological treatments were performed using the Foxm1 inhibitor RCM-1, the β-catenin inhibitor XAV-939, or a canonical Wnt pathway activator (Wnt agonist 1) (all from Selleck Chemicals, Japan).

Techniques: Isolation, Cell Culture, In Vitro, Inhibition, Control, Activation Assay