Journal: Scientific Reports

Article Title: SHP2 improves ovarian morphology and steroidogenic function in a rat PCOS model by modulating IRE1α/XBP1/NLRP3-mediated granulosa cell pyroptosis

doi: 10.1038/s41598-026-43536-2

Figure Lengend Snippet: A . Boxplot for data standardization evaluation. B . Principal component analysis scatter plot. C . Volcano plot of differentially expressed genes. D . Heatmap of differentially expressed genes. Ea. GO cellular component (CC) enrichment bubble plot. Eb. GO molecular function (MF) enrichment bubble plot. Ec. GO biological process (BP) enrichment bubble plot. Fa. KEGG pathway enrichment bubble plot. Fb. KEGG pathway enrichment lollipop plot. G . Correlation pie chart of key regulatory factors (IRE1α, XBP-1, p38, SP1, ZEB1, PKP3, Rb, E2F1, Cyclin, SHP2, BRD4). H . Scatter plot of E2F1-CDK1 co-regulation. I. Scatter plot of NLRP3-GSDMD pyroptosis axis.

Article Snippet: SHP2-overexpressing(SHP2-OE), ZEB1-overexpressing(ZEB1-KD), and PKP3-knockdown (PKP3-KD) lentiviruses were purchased from Yeasen Biotechnology (Shanghai) Co., Ltd. Hematoxylin-eosin (HE) staining kit and RIPA lysis buffer were purchased from Beyotime Biotechnology (Shanghai).

Techniques:

Journal: Scientific Reports

Article Title: SHP2 improves ovarian morphology and steroidogenic function in a rat PCOS model by modulating IRE1α/XBP1/NLRP3-mediated granulosa cell pyroptosis

doi: 10.1038/s41598-026-43536-2

Figure Lengend Snippet: SHP2 Alleviates PCOS Phenotypes via IRE1α/XBP1/NLRP3 and ZEB1/PKP3-Mediated Regulation. (A) Western blot detection of SHP2-regulated IRE1α/XBP1/ZEB1/PKP3 protein pathways. (B) Quantitative analysis of protein expression levels.* p < 0.05, ** p < 0.01, ns indicates P > 0.05; data are represented as the mean ± SD.

Article Snippet: SHP2-overexpressing(SHP2-OE), ZEB1-overexpressing(ZEB1-KD), and PKP3-knockdown (PKP3-KD) lentiviruses were purchased from Yeasen Biotechnology (Shanghai) Co., Ltd. Hematoxylin-eosin (HE) staining kit and RIPA lysis buffer were purchased from Beyotime Biotechnology (Shanghai).

Techniques: Western Blot, Expressing

Journal: Scientific Reports

Article Title: SHP2 improves ovarian morphology and steroidogenic function in a rat PCOS model by modulating IRE1α/XBP1/NLRP3-mediated granulosa cell pyroptosis

doi: 10.1038/s41598-026-43536-2

Figure Lengend Snippet: SHP2 improves ovarian morphology and steroidogenic function in a rat PCOS model by regulating the IRE1α/XBP1/NLRP3 and ZEB1/PKP3 signaling pathways, thereby influencing granulosa cell pyroptosis and proliferation.

Article Snippet: SHP2-overexpressing(SHP2-OE), ZEB1-overexpressing(ZEB1-KD), and PKP3-knockdown (PKP3-KD) lentiviruses were purchased from Yeasen Biotechnology (Shanghai) Co., Ltd. Hematoxylin-eosin (HE) staining kit and RIPA lysis buffer were purchased from Beyotime Biotechnology (Shanghai).

Techniques: Protein-Protein interactions

Journal: Frontiers in Pharmacology

Article Title: BRG1 exacerbates myocardial fibrosis after myocardial infarction by interacting with ZEB1

doi: 10.3389/fphar.2026.1802700

Figure Lengend Snippet: BRG1 interacts with ZEB1 and is required to maintain ZEB1 expression. (A) Immunofluorescence staining showing the cellular distribution of BRG1 and ZEB1 in CFs. BRG1 was stained in red, and ZEB1 was stained in green. Scale bar = 20 μm. (B,C) Protein-protein interaction between BRG1 and ZEB1 in CFs was validated by co-immunoprecipitation (IP) in CFs. (B) ZEB1 co-precipitated with anti-BRG1 antibody. (C) BRG1 co-precipitated with anti-ZEB1 antibody. (D) The interaction between BRG1 and ZEB1 in CFs treated with TGF-β1 (10 ng/mL) or untreated was confirmed by IP with anti-BRG1. (E,F) ZEB1 levels in CFs transfected with siBRG1 were quantified by qRT-PCR and Western blot. *** p < 0.001 vs. NC group by a two-tailed Student’s t-test. n = 6. (G) ZEB1 protein levels in CFs transfected with siBRG1 and treated with TGF-β1 * p < 0.05, *** p < 0.001 vs. Ctl group or TGF-β1+NC group by one-way ANOVA followed by Tukey’s post hoc analysis. n = 6.

Article Snippet: For absolute specificity, ZEB1 (Wanleibio, China, 1:1,000), Ppp2r1a (ABclonal, China, 1:1,000), phospho-Smad3 (Affinity, China, 1:500), and total Smad3 (Affinity, China, 1:500) were probed on separate membranes.

Techniques: Expressing, Immunofluorescence, Staining, Immunoprecipitation, Transfection, Quantitative RT-PCR, Western Blot, Two Tailed Test

Journal: Frontiers in Pharmacology

Article Title: BRG1 exacerbates myocardial fibrosis after myocardial infarction by interacting with ZEB1

doi: 10.3389/fphar.2026.1802700

Figure Lengend Snippet: The effects of ZEB1 knockdown in CFs. (A) Western blot analysis of ZEB1, Col-I and FN1 protein expression following ZEB1 siRNAs transfection in CFs. *** p < 0.001 vs. NC group by a two-tailed Student’s t-test. n = 6. (B) Protein levels of ZEB1, Col-I, and FN1 in CFs transfected with siZEB1 and treated with TGF-β1. * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. Ctl group or TGF-β1+NC group by two-way ANOVA followed by Tukey’s post hoc analysis. n = 6. (C) EdU fluorescence assay assessing proliferation of CFs transfected with siZEB1 and treated with TGF-β1. Scale bar = 50 μm *** p < 0.001 vs. Ctl group or TGF-β1+NC group by one-way ANOVA followed by Tukey’s post hoc analysis. n = 6. (D) Wound healing assay assessing migration of CFs transfected with siZEB1 and treated with TGF-β1. Scale bar = 100 μm * p < 0.05, *** p < 0.001 vs. Ctl group or TGF-β1+NC group by one-way ANOVA followed by Tukey’s post hoc analysis. n = 6.

Article Snippet: For absolute specificity, ZEB1 (Wanleibio, China, 1:1,000), Ppp2r1a (ABclonal, China, 1:1,000), phospho-Smad3 (Affinity, China, 1:500), and total Smad3 (Affinity, China, 1:500) were probed on separate membranes.

Techniques: Knockdown, Western Blot, Expressing, Transfection, Two Tailed Test, Fluorescence, Wound Healing Assay, Migration

Journal: Frontiers in Pharmacology

Article Title: BRG1 exacerbates myocardial fibrosis after myocardial infarction by interacting with ZEB1

doi: 10.3389/fphar.2026.1802700

Figure Lengend Snippet: ZEB1 regulated the effects of BRG1 on cardiac fibrosis. (A) Western blot analysis of ZEB1, Col-I and FN1 protein levels in CFs following co-transfection with BRG1 overexpression plasmids and siZEB1. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. Ctl group or BRG1+NC group by two-way ANOVA followed by Tukey’s post hoc analysis. n = 6. (B) Proliferative activity of CFs co-transfected with BRG1 and siZEB1 was assessed by EdU staining. Scale bar = 50 μm *** p < 0.001 vs. Ctl group or BRG1+NC group by one-way ANOVA followed by Tukey’s post hoc analysis. n = 6. (C) Cell migration was evaluated in CFs co-transfectied with BRG1 and siZEB1 by wound healing. Scale bar = 100 μm * p < 0.05, *** p < 0.001 vs. Ctl group or BRG1+NC group by one-way ANOVA followed by Tukey’s post hoc analysis. n = 6.

Article Snippet: For absolute specificity, ZEB1 (Wanleibio, China, 1:1,000), Ppp2r1a (ABclonal, China, 1:1,000), phospho-Smad3 (Affinity, China, 1:500), and total Smad3 (Affinity, China, 1:500) were probed on separate membranes.

Techniques: Western Blot, Cotransfection, Over Expression, Activity Assay, Transfection, Staining, Migration

Journal: Frontiers in Pharmacology

Article Title: BRG1 exacerbates myocardial fibrosis after myocardial infarction by interacting with ZEB1

doi: 10.3389/fphar.2026.1802700

Figure Lengend Snippet: BRG1 promoted Smad3 phosphorylation by regulating Ppp2r1a transcription. (A) Western blot analysis of p-Smad3 and t-Smad3 protein levels in CFs following co-transfection with BRG1 and siZEB1. *** p < 0.001 vs. Ctl group or BRG1+NC group by one-way ANOVA followed by Tukey’s post hoc analysis. n = 6. (B) Immunofluorescence staining reveals the nuclear translocation of Smad3 in CFs. Smad3 (green) and nuclei (DAPI, blue) were labeled. Scale bar = 25 μm. (C,D) Western blot analysis of Col-I and FN1 protein levels in CFs treated with OA following co-transfection with BRG1 overexpression plasmids and siZEB1. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. indicated groups by one-way ANOVA followed by Tukey’s post hoc analysis. n = 6. (E) In primary mouse fibroblasts with siBRG1, qRT-PCR was employed to evaluate PP2A subunit ( Ppp2ca , Ppp2cb , Ppp2r1a , Ppp2r1b , Ppp2r2a , Ppp2r2b , Ppp2r2c , Ppp2r2d , Ppp2r3a ) mRNA expression. ** p < 0.01 vs. NC group by a two-tailed Student’s t-test. n = 4. (F) Luciferase reporter assay of Ppp2r1a promoter activity in CFs transfected with siZEB1. *** p < 0.001 vs. NC group. (G) Luciferase reporter assay of Ppp2r1a promoter activity in CFs transfected with siBRG1. *** p < 0.001 vs. NC group. (H) Luciferase reporter assay of Ppp2r1a promoter activity in CFs co-transfected with BRG1 overexpression plasmid and siZEB1 *** p < 0.001 vs. Ctl group or BRG1+NC group by one-way ANOVA followed by Tukey’s post hoc analysis. n = 10. (I) ChIP analysis identified ZEB1 binding sites in the Ppp2r1a promoter region. (J) ChIP-qPCR analysis of BRG1 and ZEB1 binding to the Ppp2r1a promoter region in CFs. * p < 0.05 and ** p < 0.01 vs. IgG. n = 3. (K) Transfection efficiency of siPpp2r1a was detected by Western blot, *** p < 0.001 vs. NC group by a two-tailed Student’s t-test. n = 6. (L) Immunofluorescence detects nuclear translocation of Smad3 (green) in CFs, with DAPI (blue) marking nuclei. Scale bar = 25 μm. (M,N) Representative Western blotting analysis of t-Smad3 protein level in cytoplasm and nucleus in CFs. β-actin was used as the loading control for cytosolic fractions. Histone3 was used as the loading control for nuclear fractions. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. indicated groups by one-way ANOVA followed by Tukey’s post hoc analysis. n = 6.

Article Snippet: For absolute specificity, ZEB1 (Wanleibio, China, 1:1,000), Ppp2r1a (ABclonal, China, 1:1,000), phospho-Smad3 (Affinity, China, 1:500), and total Smad3 (Affinity, China, 1:500) were probed on separate membranes.

Techniques: Phospho-proteomics, Western Blot, Cotransfection, Immunofluorescence, Staining, Translocation Assay, Labeling, Over Expression, Quantitative RT-PCR, Expressing, Two Tailed Test, Luciferase, Reporter Assay, Activity Assay, Transfection, Plasmid Preparation, Binding Assay, ChIP-qPCR, Control

Journal: Frontiers in Pharmacology

Article Title: BRG1 exacerbates myocardial fibrosis after myocardial infarction by interacting with ZEB1

doi: 10.3389/fphar.2026.1802700

Figure Lengend Snippet: Schematic diagram depicting the role of BRG1 on cardiac fibrosis. Myocardial infarction upregulates BRG1, which interacts with ZEB1 to form a transcriptional repressor complex. This complex suppresses the expression of Ppp2r1a (encoding the PP2A structural subunit), leading to diminished PP2A activity. Consequently, Smad3 phosphorylation and nuclear translocation are enhanced, driving the transcription of fibrotic genes such as Col-1 and FN1, and ultimately exacerbating myocardial fibrosis.

Article Snippet: For absolute specificity, ZEB1 (Wanleibio, China, 1:1,000), Ppp2r1a (ABclonal, China, 1:1,000), phospho-Smad3 (Affinity, China, 1:500), and total Smad3 (Affinity, China, 1:500) were probed on separate membranes.

Techniques: Expressing, Activity Assay, Phospho-proteomics, Translocation Assay