Journal: Redox Biology

Article Title: Methylation reader MBD2-mediated GPX4 transcriptional repression drives ovarian granulosa cell ferroptosis in PCOS

doi: 10.1016/j.redox.2026.104034

Figure Lengend Snippet: GPX4 suppression is regulated by a repressive complex containing MBD2, MAZ, HDAC3 and NCoR. (a) Peak plot showing the ATAC-seq peak at the Gpx4 locus (Chr10: 80051488–80056439) in ovarian tissues from control (Ctrl, blue) and DHEA-treated (DHEA, red) mice. Orange boxes and asterisks denote regions with increased chromatin accessibility. (b) A heatmap displays the top six transcription factors (TFs) binding to the Gpx4 promoter region in the ATAC-seq analysis, along with the mRNA expression identified by RNA-seq analysis, and the predicted TF motifs and E-values are shown on the right. (c) Schematic representation of the Gpx4 promoter region showing the MAZ binding motif relative to the transcription start site (TSS). (Below) MAZ binding footprint enrichment at the Gpx4 locus in Ctrl (blue) and DHEA-treated (red) mice. Primary ovarian granulosa cells (GCs) were treated with 50 μM DHEA for 48 h in vitro to establish the PCOS model. (d) Western blot analysis of MAZ, NCoR and HDAC3 protein expression in DHEA-treated GCs. GAPDH served as a loading control. Blots are representative of one sample per group. Quantification was presented as means ± SEM, n = 3. ∗ P < 0.05, Student's t-test. (e) Co-immunoprecipitation (Co-IP) assay. Cell lysates were immunoprecipitated (IP) with isoform-matched immunoglobulin (Ig) or antibodies (IP Ab) to MBD2, MAZ, HDAC3, or NCoR, and then immunoprecipitants were assessed for MBD2, MAZ, HDAC3, or NCoR by western blotting reciprocally (the upper panel). The non-IP lysates (Input) were assayed for GAPDH as input controls. (f) Immunofluorescence co-staining was used to determine the expression and localization of MAZ (green), NCoR (red), and HDAC3 (magenta) within GCs. (g) Quantification of protein co-localization from the magnified region in ( f ). (h) Chromatin immunoprecipitation (ChIP) assay. DHEA-treated GCs were in presence or absence of KCC-07 (KCC, 10 μM, 48 h), and the cell lysates were immunoprecipitated with isoform-matched immunoglobulin or antibodies to MBD2, MAZ, NCoR, HDAC3, or pan-acetylated lysine (Pan-Ace), respectively. The genomic DNA (Input) and the antibody-bound DNAs were PCR-amplified with primers covering the MAZ motif on Gpx4 promoter. The PCR products of representative sample per group were analyzed on 1.5 % agarose gels. Quantitative analysis was shown on the right. Data were presented as mean ± SEM, n = 4. ∗ P < 0.05, one-way ANOVA. (i) Western blot analysis. (Left) HDAC3 and GPX4 protein expression in DHEA-treated GCs in the presence or absence of the HDAC3 inhibitor RGFP966 (RGFP, 10 μM, 48 h). (Middle) MAZ and GPX4 protein expression in GCs transfected with negative- (si-Ctrl) or MAZ-targeting (si-MAZ) siRNA, followed by treatment with or without DHEA. (Right) NCoR and GPX4 protein expression in GCs transfected with negative- (si-Ctrl) or NCoR-targeting (si-NCoR) siRNA, followed by DHEA treatment. GAPDH was as a loading control. (j) Quantifications of ( i ). Data were presented as mean ± SEM, n = 3. ∗ P < 0.05, one-way ANOVA. (k) Schematic model of Gpx4 transcriptional repression. A transcriptional repressive complex orchestrated by MBD2, MAZ, HDAC3, and NCoR binds to the hypermethylated Gpx4 promoter, leading to transcriptional suppression.

Article Snippet: A PCOS cell model was established by DHEA (50 μM, 48 h) treatment, followed by KCC-07 (10 μM, 48 h), 5-Aza (10 μM, 24 h), RGFP966 (10 μM, 48 h, HY-13909, MCE, USA), and/or Liproxstatin-1 (200 nM, 48 h, HY-12726, MCE, USA) treatments.

Techniques: Control, Binding Assay, Expressing, RNA Sequencing, In Vitro, Western Blot, Co-Immunoprecipitation Assay, Immunoprecipitation, Immunofluorescence, Staining, Chromatin Immunoprecipitation, Amplification, Transfection

Journal: International Journal of Nanomedicine

Article Title: LincRNA-p21: A Double-Edged Sword in Ethanol-Induced Liver Damage and Its Nanoparticle Solution

doi: 10.2147/IJN.S577455

Figure Lengend Snippet: ALKBH5/lincRNA-p21/FGFR2 promotes autophagy in response to ethanol-induced liver injury. ( A ) HepG2 cells were treated with/without 10 μM SAHA, 10 μM NaB, or 5 μM RGFP966 for 24 h, the expression of lincRNA-p21 was evaluated by RT-PCR. n=3. ( B ) The potential m 6 A sites in lincRNA-p21 predicted using SRAMP. ( C ) m 6 A RNA methylation levels of HepG2 cells with/without alcoholic hepatocyte injury ( P >0.05). n=6. ( D ) When HepG2 cells were exposed to ethanol, the expressions of lincRNA-p21, ALKBH5 and METTL3 were evaluated. The expressions of lincRNA-p21 and ALKBH5 were all down-regulated when HepG2 cells were exposed to 100 mM ethanol ( P <0.01, P <0.05). ( E ) The location of lincRNA-p21 in HepG2 cells demonstrated by the lincRNA-p21 FISH Probe. Scale bars: 50 μm. ( F ) The expressions of lincRNA-p21 was upregulated by ALKBH5 overexpression. n=3. ( G ) LincRNA-p21 was significantly enriched in ALKBH5-bound RNA. n=3. ( H )The methylation level of the 1237 nt site in lincRNA-p21 was evaluated using MeRIP-PCR. n=3. ( I ) Schematic diagram of ALKBH5 mediating m 6 A demethylation and upregulation of lincRNA-p21. The red downward arrow indicates decreased expression. ( J ) The mRNA and protein expression of FGFR2 was evaluated by RT-PCR and Western blot. FGFR2 protein was up-regulated by the overexpression of lincRNA-p21 (FGFR2/β-actin: 0.54 vs 0.20, P =0.001). ( K ) Fluorescence images of the autophagosomes. Scale bars: 10 μm. ( L ) Cell viability was evaluated using a CCK8 assay. n=3. Data represent mean±s.e.m. Significance was determined by two-tailed Student’s t -test. * P <0.05, ** P <0.01, *** P <0.001.

Article Snippet: The HDAC inhibitors Vorinostat (SAHA)(#HY-10221, MCE, USA), sodium butyrate (NaB)(#S1999, Selleck, USA), and HDAC3 inhibitor RGFP966 (#HY-13909, MCE, USA) were used to study the mechanism of lincRNA-p21 regulation.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Methylation, Over Expression, Western Blot, Fluorescence, CCK-8 Assay, Two Tailed Test

Journal: Frontiers in Medicine

Article Title: Investigating the causal relationship of lipid metabolism in polycystic ovary syndrome: a Mendelian randomization study on the regulatory role of 3-Hydroxybutyrate in gene expression

doi: 10.3389/fmed.2026.1747593

Figure Lengend Snippet: (A) 3-HB; (B) RGFP966. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: HDAC3-specific inhibitor RGFP966 (MCE, Catalog No. HY-13909), 3-Hydroxybutyrate (MCE, Catalog No. HY-113378).

Techniques:

Journal: Frontiers in Medicine

Article Title: Investigating the causal relationship of lipid metabolism in polycystic ovary syndrome: a Mendelian randomization study on the regulatory role of 3-Hydroxybutyrate in gene expression

doi: 10.3389/fmed.2026.1747593

Figure Lengend Snippet: Results of sensitivity analysis-forest. (A) Ratio of apolipoprotien B to apolipoprotien A1 levels; (B) 3-hydroxybutyrate levels; (C) triglyceride levels in LDL; (D) triglycerides to total lipids ratio in large LDL; (E) triglyceride in medium LDL; (F) total cholesterol levels in very large HDL; (G) triglycerides to total lipids ratio in very large HDL; (H) total lipids in very small VLDL; (I) concentration of very small VLDL particles.

Article Snippet: HDAC3-specific inhibitor RGFP966 (MCE, Catalog No. HY-13909), 3-Hydroxybutyrate (MCE, Catalog No. HY-113378).

Techniques: Concentration Assay

Journal: Frontiers in Medicine

Article Title: Investigating the causal relationship of lipid metabolism in polycystic ovary syndrome: a Mendelian randomization study on the regulatory role of 3-Hydroxybutyrate in gene expression

doi: 10.3389/fmed.2026.1747593

Figure Lengend Snippet: Results of sensitivity analysis-funnelplot. (A) Ratio of apolipoprotien B to apolipoprotien A1 levels; (B) 3-hydroxybutyrate levels; (C) triglyceride levels in LDL; (D) triglycerides to total lipids ratio in large LDL; (E) triglyceride in medium LDL; (F) total cholesterol levels in very large HDL; (G) triglycerides to total lipids ratio in very large HDL; (H) total lipids in very small VLDL; (I) concentration of very small VLDL particles.

Article Snippet: HDAC3-specific inhibitor RGFP966 (MCE, Catalog No. HY-13909), 3-Hydroxybutyrate (MCE, Catalog No. HY-113378).

Techniques: Concentration Assay