Journal: Redox Biology

Article Title: ROS-activated CD147-type I interferon signaling axis drives vascular smooth muscle cell fate transition and abdominal aortic aneurysm progression

doi: 10.1016/j.redox.2025.103780

Figure Lengend Snippet: CD147 interacts with STAT1/STAT2 complex to activate IFN-I signaling in VSMCs. (A) Schematic workflow of the CD147 protein interactome analysis. Senescent VSMCs were infected with adenovirus expressing FLAG-CD147. Cell lysates were immunoprecipitated using anti-FLAG beads and subjected to mass spectrometry for protein identification. (B) Five unique peptides from the STAT1/STAT2 complex were identified ( left ), and a representative LC-MS/MS spectrum is shown ( right ). (C) Western blot analysis showing the association of FLAG-CD147 with the STAT1/STAT2 complex in VSMCs. FLAG-CD147 was immunoprecipitated with anti-FLAG antibody from VSMC lysates, followed by Western blotting. (D) Predicted structural model of the CD147/STAT1/STAT2 complex generated by AlphaFold3. (E) VSMCs were infected with adenovirus expressing FLAG-CD147 and either STAT2-targeting siRNA or a negative control (si-NC, scrambled siRNA). Lysates were immunoprecipitated with anti-FLAG antibody, followed by Western blot analysis. (F) Western blot analysis of STAT2, IRF7, IFNα, IFNβ, P21, α-SMA, SM22, CNN1 and MYH11 protein levels in VSMCs transfected with CD147-OE adenovirus and either STAT2-targeting siRNA or negative control (si-NC, scrambled siRNA), followed by exposure to PBS or H 2 O 2 (100 μM). (G) Left: Representative immunofluorescence images showing IRF7, P16, P21 expression, and SA-β-gal staining in VSMCs transfected with CD147-OE adenovirus and either STAT2-targeting siRNA or negative control, followed by exposure to PBS or H 2 O 2 (100 μM). The slides were co-stained with SMC marker α-SMA and DAPI. Right: Quantification of IRF7, P16, P21 protein expression levels, and SA-β-gal positive areas in the indicated groups (n = 10). ∗∗∗∗ P < 0.0001.

Article Snippet: Small interfering RNAs (siRNAs) against rat STAT2 and a scrambled siRNA were designed and synthesized by OBiO Technology (Shanghai, China).

Techniques: Infection, Expressing, Immunoprecipitation, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Western Blot, Generated, Negative Control, Transfection, Immunofluorescence, Staining, Marker

Journal: Redox Biology

Article Title: ROS-activated CD147-type I interferon signaling axis drives vascular smooth muscle cell fate transition and abdominal aortic aneurysm progression

doi: 10.1016/j.redox.2025.103780

Figure Lengend Snippet: CD147 knockout significantly alleviated AngII-induced VSMC phenotypic transition, IFN-I response, and MMP activation. (A–B) Representative Western blot and quantification of STAT2, IRF7, α-SMA, SM22, CNN1, and MYH11 protein levels in abdominal aortas in the indicated groups of mice on day 28 following AngII infusion. ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001. (C – D) Representative immunohistochemistry images and quantification of typical matrix-degrading enzyme MMP2 and inflammatory markers (IL-6, TNF-α) protein expression levels in aneurysmal tissues from the indicated groups of mice on day 28 following AngII infusion (n = 5). ∗ P < 0.05. (E – F) Representative immunofluorescence images and quantification of fluorescence intensity for protein expression of IFN-I components (IRF7, IFNβ) and senescence marker (P21) in aneurysmal tissues from the indicated groups of mice on day 28 following AngII infusion. ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001. (G) Representative images and quantification of active MMP in aneurysmal tissues from the indicated groups of mice on day 28 following AngII infusion (n = 6). ∗∗∗ P < 0.001. (H) MMP activity measured by gelatin zymography in isolated VSMCs treated with AngII (1 μM). ∗∗ P < 0.01.

Article Snippet: Small interfering RNAs (siRNAs) against rat STAT2 and a scrambled siRNA were designed and synthesized by OBiO Technology (Shanghai, China).

Techniques: Knock-Out, Activation Assay, Western Blot, Immunohistochemistry, Expressing, Immunofluorescence, Fluorescence, Marker, Activity Assay, Zymography, Isolation

Journal: Redox Biology

Article Title: ROS-activated CD147-type I interferon signaling axis drives vascular smooth muscle cell fate transition and abdominal aortic aneurysm progression

doi: 10.1016/j.redox.2025.103780

Figure Lengend Snippet: Administration of the CD147 natural inhibitor Myricetin attenuates AngII-induced VSMC phenotypic transition, IFN-I response, and MMP activation. (A–B) Western blot and quantification of STAT2, IRF7, α-SMA, SM22, CNN1, and MYH11 protein levels in abdominal aortas in the indicated groups of mice on day 28 following AngII infusion. ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001. (C – D) Representative immunohistochemistry images and quantification of typical matrix-degrading enzyme MMP2 and inflammatory markers (IL-6, TNF-α) protein expression levels in aneurysmal tissues from the indicated groups of mice on day 28 following AngII infusion (n = 5). ∗ P < 0.05; ∗∗∗ P < 0.001. (E – F) Representative immunofluorescence images and quantification of fluorescence intensity for protein expression of IFN-I components (IFNα, IFNβ) and senescence marker (P21) in aneurysmal tissues from the indicated groups of mice on day 28 following AngII infusion (n = 5). ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001. (G) Representative images and quantification of active MMP in aneurysmal tissues from the indicated groups of mice on day 28 following AngII infusion (n = 6). ∗ P < 0.05. (H) MMP activity measured by gelatin zymography in isolated VSMCs treated with AngII (1 μM). ∗∗∗∗ P < 0.0001.

Article Snippet: Small interfering RNAs (siRNAs) against rat STAT2 and a scrambled siRNA were designed and synthesized by OBiO Technology (Shanghai, China).

Techniques: Activation Assay, Western Blot, Immunohistochemistry, Expressing, Immunofluorescence, Fluorescence, Marker, Activity Assay, Zymography, Isolation

Journal: International Journal of Biological Sciences

Article Title: Glycine Decarboxylase Regulates Renal Carcinoma Progression via Interferon Stimulated Gene Factor 3-Mediated Pathway

doi: 10.7150/ijbs.104458

Figure Lengend Snippet: GLDC regulates RCC cell progression via ISGF3 pathway. (A) Western blot of GLDC, IRF9, STAT2, STAT1, SETD2, GAPDH (loading control), H3K36me3, and H3 (loading control for H3K36me3) in indicated cells. To confirm in Caki-2 cells, we suppressed the expression of GLDC in Caki-2 cells with three shRNA constructs (shGLDC #1, shGLDC #4 and shGLDC #5), and observed the expression of indicated proteins. (B) Quantitative PCR (qPCR) analysis of the ISGs: OSA1, IFIT1, IFI44, IFI44L, CCL5, MX1, and IFI27 of indicated cells. Data are shown as the means ± SD ( n =3). (C) Western blot analysis of indicated proteins in ACHN CTL and shGLDC cells transfected with siRNA of negative control (siNC), STAT2 and/ or IRF9 in ACHN knock-downed GLDC cells compared to control (siNC)-siRNA cells. The cellular growth of indicated cells was on the right side ( n =3). (D) Colony formation of indicated cells. Quantification of staining intensity was performed at a wavelength of 570 nm ( n =3). (E) qPCR analysis of IFN-α in ACHN CTL and shGLDC treated with Poly (I:C) (10 µg) or transfected with Poly (I:C) (1 µg) for 24 h. Data are shown as the means ± SD ( n =3). p values were calculated using two-tailed unpaired Student t-test. * p < 0.05, ** p < 0.01, and *** p < 0.001. CTL, control; shGLDC, knockdown of GLDC; EV, corresponding control for OE GLDC; OE GLDC, overexpression of GLDC.

Article Snippet: Short interfering RNAs (siRNAs) for STAT2 and IRF9 came from Bioneer, Republic of Korea.

Techniques: Western Blot, Control, Expressing, shRNA, Construct, Real-time Polymerase Chain Reaction, Transfection, Negative Control, Staining, Two Tailed Test, Knockdown, Over Expression

Journal: International Journal of Biological Sciences

Article Title: Glycine Decarboxylase Regulates Renal Carcinoma Progression via Interferon Stimulated Gene Factor 3-Mediated Pathway

doi: 10.7150/ijbs.104458

Figure Lengend Snippet: Suppression of GLDC inhibits RCC progression in vivo . (A) Images of xenograft tumor derived from injection of ACHN control (CTL) and GLDC knock-downed cells (shGLDC) ( n =7) and A498 control (EV) and GLDC over-expressed cells (OE GLDC) ( n =4). (B) Quantification of tumor volume and tumor weight from ACHN CTL and shGLDC cells. (C) Quantification of tumor volume and tumor weight from A498 EV and OE GLDC cells. (D) Immunohistochemistry staining of GLDC, STAT2, and Ki-67 in tumors injected with ACHN CTL and shGLDC cells. (E) Western blot of GLDC, IRF9, STAT2, STAT1, and GAPDH (loading control) in tumors injected with ACHN CTL and shGLDC cells. (F) Western blot of GLDC and actin (loading control) of tumor and non-tumor tissues from patients with high-risk RCC. p values were calculated using two-tailed unpaired Student t -test. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: Short interfering RNAs (siRNAs) for STAT2 and IRF9 came from Bioneer, Republic of Korea.

Techniques: In Vivo, Derivative Assay, Injection, Control, Immunohistochemistry, Staining, Western Blot, Two Tailed Test

Journal: Cancers

Article Title: Enhanced IFNα Signaling Promotes Ligand-Independent Activation of ERα to Promote Aromatase Inhibitor Resistance in Breast Cancer

doi: 10.3390/cancers13205130

Figure Lengend Snippet: STAT1 and STAT2 expression affect ERα and ER-regulated gene expression in AI-resistant breast cancer cells. ( A ) MCF-7 and MCF-7:5C cells were transiently transfected for 48 h with siRNA against STAT1 or STAT2 and immunoblotted for the proteins indicated. ( B ) MCF-7 and MCF-7:5C cells were transiently transfected for the ERE reporter construct and siRNA against STAT1 or STAT2 for 48 h. Luciferase activity was then read. ( C , D ) MCF-7 and MCF-7:5C cells were transfected with siCon, siSTAT1, or siSTAT2 (as indicated) and analyzed by RT-PCR. * p < 0.05, ** p < 0.01 and *** p < 0.001.

Article Snippet: T47D, MCF-7, and MCF-7:5C cells (1 × 10 5 ) were transiently transfected with siRNAs for ERα (Santa Cruz Biotechnology, Dallas, TX, USA, Cat#sc-29305), STAT1 (Santa Cruz Biotechnology, Cat#sc-44123), and STAT2 (Santa Cruz Biotechnology, Cat#sc-29492) or a scrambled negative control (Santa Cruz Biotechnology, Cat#sc-37007).

Techniques: Expressing, Gene Expression, Transfection, Construct, Luciferase, Activity Assay, Reverse Transcription Polymerase Chain Reaction

Journal: Cancers

Article Title: Enhanced IFNα Signaling Promotes Ligand-Independent Activation of ERα to Promote Aromatase Inhibitor Resistance in Breast Cancer

doi: 10.3390/cancers13205130

Figure Lengend Snippet: STAT1 interacts with ERα through in silico and in vitro analysis in AI-resistant breast cancer cells. ( A ) X-ray crystal structures of STAT1 and ERα were downloaded from PDB and prepared for docking by removing ligands, water molecules and extra chain of amino acids. Chain A was selected for both proteins. The proteins were further prepared using MGL tools. The final prepared protein structures were uploaded to the GRAMM-X protein docking server for checking interactions. The final output file was analyzed using PYMOL program. ( B ) T47D, MCF-7, and MCF-7:5C cells were immunoprecipitated with anti-ERα or rabbit IgG and immunoblotted for STAT1, STAT2, IRF9 and ERα. ( C ) All cell lines were grown on coverslips and processed with the Duolink ® PLA Fluorescence with ERα and STAT1 antibodies.

Article Snippet: T47D, MCF-7, and MCF-7:5C cells (1 × 10 5 ) were transiently transfected with siRNAs for ERα (Santa Cruz Biotechnology, Dallas, TX, USA, Cat#sc-29305), STAT1 (Santa Cruz Biotechnology, Cat#sc-44123), and STAT2 (Santa Cruz Biotechnology, Cat#sc-29492) or a scrambled negative control (Santa Cruz Biotechnology, Cat#sc-37007).

Techniques: In Silico, In Vitro, Immunoprecipitation, Fluorescence