Journal: iScience

Article Title: RSK1-SRF signaling axis drives fibroblast activation and pulmonary fibrosis: Genetic causality and therapeutic targeting

doi: 10.1016/j.isci.2026.115495

Figure Lengend Snippet: Inhibition of SRF prevents TGF-β-induced fibroblast activation and ECM deposition (A) Immunofluorescence co-staining of p -SRF (green) with F4/80, E-Cad, and FN (red) in saline- and BLM-treated mouse lungs; nuclei are stained with DAPI (blue). Scale bars, 100 μm. (B) Immunoblot analysis of p -SRF and total SRF in HFL-1 cells treated with TGF-β (10 ng/mL) with or without CCG-1423 (10 μM) for 72 h. (C) Densitometric quantification of (B); GAPDH serves as the loading control. (D) Representative immunofluorescence images of p -SRF in HFL-1 cells treated as in (B). Scale bars, 20 μm. (E) Mean fluorescence intensity quantification of (D). (F) Immunoblot analysis of FN and α-SMA in HFL-1 cells treated as in (B). (G) Densitometric quantification of (F); GAPDH serves as the loading control. (H–J) Representative immunofluorescence images of FN (H), collagen II (I), and α-SMA (J) in HFL-1 cells treated as in (B). Scale bars, 20 μm. (K–M) Mean fluorescence intensity quantification of (H–J), respectively. Data are presented as the mean ± SD. For cell-based assays, n denotes independent biological replicates (independent cell culture batches processed in separate experiments; n = 3). Image quantification was performed as described in ; technical sampling was not counted toward n . Statistical tests are described in . ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; # p ≥ 0.05.

Article Snippet: CCG-1423 , MedChemExpress , Cat#HY-13991.

Techniques: Inhibition, Activation Assay, Immunofluorescence, Staining, Saline, Western Blot, Control, Fluorescence, Cell Culture, Sampling

Journal: iScience

Article Title: RSK1-SRF signaling axis drives fibroblast activation and pulmonary fibrosis: Genetic causality and therapeutic targeting

doi: 10.1016/j.isci.2026.115495

Figure Lengend Snippet: Superior anti-fibrotic efficacy of targeting the RSK1-SRF axis in pulmonary fibrosis (A) Representative immunohistochemistry (IHC) staining images of p -SRF in control, BLM-treated, and BLM + CCG-1423 (0.15 mg/kg, daily for 7 days) mouse lungs. Scale bars, 100 μm. (B) Quantification of p -SRF IHC staining shown as positive area (%). (C) Immunoblot analysis of p -SRF and total SRF in the indicated groups. (D) Densitometric quantification of (C); β-actin serves as the loading control. (E) Immunoblot analysis of FN, collagen II, vimentin, and α-SMA in the indicated groups. (F) Densitometric quantification of (E); β-actin serves as the loading control. (G) Immunoblot analysis of p -RSK1 and total RSK1 in control, BLM-treated, and BLM + BID-1870 (0.5 mg/kg, daily for 7 days) mouse lungs. (H) Densitometric quantification of (G); β-actin serves as the loading control. (I) Representative IHC staining images of p -RSK1 in the indicated groups. Scale bars, 100 μm. (J) Quantification of p -RSK1 IHC staining shown as positive area (%). (K) Immunoblot analysis of FN, collagen II, vimentin, and α-SMA in the indicated groups. (L) Densitometric quantification of (K); β-actin serves as the loading control. (M) Immunoblot analysis of p -SRF and total SRF in the indicated groups. (N) Densitometric quantification of (M); β-actin serves as the loading control. (O) Representative IHC staining images of p -SRF in the indicated groups. Scale bars, 100 μm. (P) Quantification of p -SRF IHC staining shown as positive area (%). (Q–S) Histological analyses of lung fibrosis by H&E, Masson’s trichrome, and Sirius Red staining in the indicated groups. Scale bars, 100 μm. Data are presented as the mean ± SD. For tissue-based assays, n denotes independent animals ( n = 6 per group). Image quantification was performed as described in ; technical sampling was not counted toward n . Statistical tests are described in . ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; # p ≥ 0.05.

Article Snippet: CCG-1423 , MedChemExpress , Cat#HY-13991.

Techniques: Immunohistochemistry, Control, Western Blot, Staining, Sampling

Journal: iScience

Article Title: RSK1-SRF signaling axis drives fibroblast activation and pulmonary fibrosis: Genetic causality and therapeutic targeting

doi: 10.1016/j.isci.2026.115495

Figure Lengend Snippet: Inhibition of SRF prevents TGF-β-induced fibroblast activation and ECM deposition (A) Immunofluorescence co-staining of p -SRF (green) with F4/80, E-Cad, and FN (red) in saline- and BLM-treated mouse lungs; nuclei are stained with DAPI (blue). Scale bars, 100 μm. (B) Immunoblot analysis of p -SRF and total SRF in HFL-1 cells treated with TGF-β (10 ng/mL) with or without CCG-1423 (10 μM) for 72 h. (C) Densitometric quantification of (B); GAPDH serves as the loading control. (D) Representative immunofluorescence images of p -SRF in HFL-1 cells treated as in (B). Scale bars, 20 μm. (E) Mean fluorescence intensity quantification of (D). (F) Immunoblot analysis of FN and α-SMA in HFL-1 cells treated as in (B). (G) Densitometric quantification of (F); GAPDH serves as the loading control. (H–J) Representative immunofluorescence images of FN (H), collagen II (I), and α-SMA (J) in HFL-1 cells treated as in (B). Scale bars, 20 μm. (K–M) Mean fluorescence intensity quantification of (H–J), respectively. Data are presented as the mean ± SD. For cell-based assays, n denotes independent biological replicates (independent cell culture batches processed in separate experiments; n = 3). Image quantification was performed as described in ; technical sampling was not counted toward n . Statistical tests are described in . ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; # p ≥ 0.05.

Article Snippet: CCG-1423 (MedChemExpress, Cat# HY-13991) stock: 10 mg CCG-1423 was dissolved in 1.0995 mL DMSO to obtain a 20 mM stock solution.

Techniques: Inhibition, Activation Assay, Immunofluorescence, Staining, Saline, Western Blot, Control, Fluorescence, Cell Culture, Sampling

Journal: iScience

Article Title: RSK1-SRF signaling axis drives fibroblast activation and pulmonary fibrosis: Genetic causality and therapeutic targeting

doi: 10.1016/j.isci.2026.115495

Figure Lengend Snippet: Superior anti-fibrotic efficacy of targeting the RSK1-SRF axis in pulmonary fibrosis (A) Representative immunohistochemistry (IHC) staining images of p -SRF in control, BLM-treated, and BLM + CCG-1423 (0.15 mg/kg, daily for 7 days) mouse lungs. Scale bars, 100 μm. (B) Quantification of p -SRF IHC staining shown as positive area (%). (C) Immunoblot analysis of p -SRF and total SRF in the indicated groups. (D) Densitometric quantification of (C); β-actin serves as the loading control. (E) Immunoblot analysis of FN, collagen II, vimentin, and α-SMA in the indicated groups. (F) Densitometric quantification of (E); β-actin serves as the loading control. (G) Immunoblot analysis of p -RSK1 and total RSK1 in control, BLM-treated, and BLM + BID-1870 (0.5 mg/kg, daily for 7 days) mouse lungs. (H) Densitometric quantification of (G); β-actin serves as the loading control. (I) Representative IHC staining images of p -RSK1 in the indicated groups. Scale bars, 100 μm. (J) Quantification of p -RSK1 IHC staining shown as positive area (%). (K) Immunoblot analysis of FN, collagen II, vimentin, and α-SMA in the indicated groups. (L) Densitometric quantification of (K); β-actin serves as the loading control. (M) Immunoblot analysis of p -SRF and total SRF in the indicated groups. (N) Densitometric quantification of (M); β-actin serves as the loading control. (O) Representative IHC staining images of p -SRF in the indicated groups. Scale bars, 100 μm. (P) Quantification of p -SRF IHC staining shown as positive area (%). (Q–S) Histological analyses of lung fibrosis by H&E, Masson’s trichrome, and Sirius Red staining in the indicated groups. Scale bars, 100 μm. Data are presented as the mean ± SD. For tissue-based assays, n denotes independent animals ( n = 6 per group). Image quantification was performed as described in ; technical sampling was not counted toward n . Statistical tests are described in . ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; # p ≥ 0.05.

Article Snippet: CCG-1423 (MedChemExpress, Cat# HY-13991) stock: 10 mg CCG-1423 was dissolved in 1.0995 mL DMSO to obtain a 20 mM stock solution.

Techniques: Immunohistochemistry, Control, Western Blot, Staining, Sampling

Journal: bioRxiv

Article Title: Myofibroblast lineage mapping and inhibiting subretinal fibrosis by targeting SMAD3 and MRTF pathways via microRNA-24 functional study

doi: 10.64898/2026.03.03.709397

Figure Lengend Snippet: a Representative immunostaining images showing repressed expression of Fibronectin and F-CHP staining by SMAD3 inhibitor (SIS3) at 10 μM or MRTF-A inhibitor (CCG-203971) at 20 μM, individually or in combination, in TGF-β2-treated ARPE-19 cells. B Quantification of ( a ). n=3. c Experimental timeline for evaluation of dual inhibition of SMAD3 and MRTF activity in the laser-induced subretinal fibrosis model. Representative three-dimensional reconstructions images showing RPE/choroid flatmount staining for F-CHP (green), Fibronectin (red), and α-SMA (magenta) are shown, with individual channels displayed below. d Quantification of ( c ). n = 6 mice per group, with 3 laser spots per eye. Eyes with hemorrhage after laser photocoagulation were excluded from analysis. *p < 0.05; * *p < 0.01; ***p < 0.001; ****p < 0.0001; ns, not significant.

Article Snippet: Beginning 7 days after laser photocoagulation, mice received daily intraperitoneal injections of the MRTF-A inhibitor CCG-203971 (MedChemExpress, HY-108361)(100 mg/kg), the SMAD3 inhibitor SIS3 (Sigma, S0447) (2.5 mg/kg), or a combination of both compounds for an additional 2 weeks.

Techniques: Immunostaining, Expressing, Staining, Inhibition, Activity Assay

Journal: bioRxiv

Article Title: Myofibroblast lineage mapping and inhibiting subretinal fibrosis by targeting SMAD3 and MRTF pathways via microRNA-24 functional study

doi: 10.64898/2026.03.03.709397

Figure Lengend Snippet: a Representative immunostaining images showing repressed expression of Fibronectin and F-CHP staining by SMAD3 inhibitor (SIS3) at 10 μM or MRTF-A inhibitor (CCG-203971) at 20 μM, individually or in combination, in TGF-β2-treated ARPE-19 cells. B Quantification of ( a ). n=3. c Experimental timeline for evaluation of dual inhibition of SMAD3 and MRTF activity in the laser-induced subretinal fibrosis model. Representative three-dimensional reconstructions images showing RPE/choroid flatmount staining for F-CHP (green), Fibronectin (red), and α-SMA (magenta) are shown, with individual channels displayed below. d Quantification of ( c ). n = 6 mice per group, with 3 laser spots per eye. Eyes with hemorrhage after laser photocoagulation were excluded from analysis. *p < 0.05; * *p < 0.01; ***p < 0.001; ****p < 0.0001; ns, not significant.

Article Snippet: CCG-203971 (MedChemExpress, HY-108361) is a MTRF inhibitor functioning by blocking MRTF nuclear accumulation.

Techniques: Immunostaining, Expressing, Staining, Inhibition, Activity Assay

Journal: Redox Biology

Article Title: Myo1f regulates monocyte adhesion and contributes to atherosclerosis via MRTFA-dependent ITGB2 expression

doi: 10.1016/j.redox.2026.104049

Figure Lengend Snippet: Myo1f promotes ITGB2 transcription through MRTFA. A, The adhesion of THP-1 cells to HUVECs after treatment with MRTFA knockdown lentivirus was observed under a fluorescence microscope (scale bar, 50 μm). Adherent cell numbers were calculated from 3 biological replicate images, and data are shown as fold change (ratio to shCtrl). B, Western blot analysis of MRTFA protein levels in THP-1 cells administered or not administered oxLDL (n = 3). C, Confirm the effectiveness of MRTFA knockdown lentivirus by western blotting (n = 3). D, Representative Western blot and quantification of ITGB2 expression after THP-1 knockdown of MRTFA with or without exposure to oxLDL (n = 3). E, Detection of Itgb2 mRNA levels of THP-1 under oxLDL stimulation by qPCR after elimination or not elimination of MRTFA (n = 5). F, Western blotting analysis of ITGB2 expression after gradient increase of MRTFA inhibitor CCG-1423 under oxLDL stimulation (n = 3). G, Representative confocal images of ITGB2 (red) with or without MRTFA knockdown and THP-1 stimulation with oxLDL (scale bar, 20 μm; n = 3). H, Western blot analysis of ITGB2 expression in oxLDL-stimulated THP-1 cells after overexpression of Myo1f with or without elimination of MRTFA (n = 3). I, Nuclear translocation of MRTFA following elimination of Myo1f based on oxLDL stimulation by dissociating cytoplasmic and nuclear analysis (n = 3). Data were presented as the mean ± SEM. Student's t-test (unpaired, two-tailed) for B, E and G . One-way ANOVA followed by Tukey's multiple comparisons test for A, C, D, F, H and I . Each P value is displayed in the image.

Article Snippet: Eight-week-old Apoe −/− mice were randomly divided into groups and simultaneously intraperitoneally injected with the MRTFA inhibitor (CCG-1423, 1 mg/kg, HY-13991, MCE) or vehicle and were given high-cholesterol feed for 8 weeks.

Techniques: Knockdown, Fluorescence, Microscopy, Western Blot, Expressing, Over Expression, Translocation Assay, Two Tailed Test

Journal: Redox Biology

Article Title: Myo1f regulates monocyte adhesion and contributes to atherosclerosis via MRTFA-dependent ITGB2 expression

doi: 10.1016/j.redox.2026.104049

Figure Lengend Snippet: Cytoskeletal polymerization promotes ITGB2 transcription and MRTFA nuclear translocation. A, Western blot analysis of ITGB2 expression after increasing gradient of G-actin polymerization inhibitor LAT-A under oxLDL stimulation (n = 3). B, The adhesion of THP-1 cells to HUVECs after treatment with LAT-A (2 μM) was observed under a fluorescence microscope (scale bar, 50 μm). Adherent cell numbers were calculated from 3 biological replicate images, and data are shown as fold change (ratio to DMSO). C, After overexpressing Myo1f and treating THP-1 cells with or without LAT-A, Western blot analysis was performed on ITGB2 expression under oxLDL stimulation (n = 3). D, Nuclear translocation of MRTFA after LAT-A treatment based on oxLDL stimulation by dissociated cytoplasmic and nuclear analysis (n = 3). E, Representative colocalization immunofluorescence micrographs of phalloidin-labeled F-actin (red) and Myo1f (green) staining in THP-1 cells. The scan line graph represents the F-actin and Myo1f staining intensity along the white straight line (scale bar, 20 μm; n = 3). F, By isolating F-actin and G-actin in THP-1 cells exposed to oxLDL with or without depletion of Myo1f, and quantifying their ratios by immunoblotting (n = 3). G, Effects of knockdown of Myo1f on MRTFA and actin binding in THP-1 cells. MRTFA antibody was used to co-immunoprecipitate actin in THP-1 cells with or without Myo1f knockdown treatment and stimulation with oxLDL (n = 3). Data were presented as the mean ± SEM. Student's t-test (unpaired, two-tailed) for F and G . One-way ANOVA followed by Tukey's multiple comparisons test for A-D . Each P value is displayed in the image.

Article Snippet: Eight-week-old Apoe −/− mice were randomly divided into groups and simultaneously intraperitoneally injected with the MRTFA inhibitor (CCG-1423, 1 mg/kg, HY-13991, MCE) or vehicle and were given high-cholesterol feed for 8 weeks.

Techniques: Translocation Assay, Western Blot, Expressing, Fluorescence, Microscopy, Immunofluorescence, Labeling, Staining, Knockdown, Binding Assay, Two Tailed Test

Journal: Redox Biology

Article Title: Myo1f regulates monocyte adhesion and contributes to atherosclerosis via MRTFA-dependent ITGB2 expression

doi: 10.1016/j.redox.2026.104049

Figure Lengend Snippet: Inhibiting MRTFA alleviates atherosclerosis. A, Schematic diagram of the intervention process by intraperitoneally injecting the MRTFA inhibitor CCG-1423 into Apoe −/− mice. B, Representative images of en face Oil Red O-stained aortas of Apoe −/− and Apoe −/− mice intraperitoneally injected with CCG-1423 are shown (n = 7). C, Representative images of HE and Oil Red O staining of cross-sections of aortic roots in mice with the indicated groups (scale bar, 200 μm; n = 7). D-F, Body weight and total plasma cholesterol and LDL-c levels in specific groups of mice (n = 7). G-I, The serum ALT, AST and Cr levels in specific groups of mice (n = 7). J, Western blot analysis of ITGB2 levels in mouse aortic plaques of mice in indicated groups (n = 7). K, Proposed mechanism of Myo1f in regulating monocyte adhesion and atherosclerosis progression. Data were presented as the mean ± SEM. Student's t-test (unpaired, two-tailed) for B-J . Each P value is displayed in the image.

Article Snippet: Eight-week-old Apoe −/− mice were randomly divided into groups and simultaneously intraperitoneally injected with the MRTFA inhibitor (CCG-1423, 1 mg/kg, HY-13991, MCE) or vehicle and were given high-cholesterol feed for 8 weeks.

Techniques: Staining, Injection, Clinical Proteomics, Western Blot, Two Tailed Test