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 knockout in granulosa cells aggregates ovarian ferroptosis. (a) Generation of granulosa cell-specific Gpx4 knockout mice ( Gpx4 GC−/− ) by crossing Gpx4 fl/fl with Cyp19α1 -Cre mice. Gpx4 locus in wild-type mice were represented by boxes (exons 1–7). The positions of Cyp19a1-Cre genotyping PCR primers F2 and R2 (arrows), and the loxP sites (blue triangles) in Gpx4 fl/fl mice and the genotyping PCR primers F1 and R1 (arrows) were depicted. (b) Genotyping of mice by PCR. Agarose gel electrophoresis shows PCR products from WT, Gpx4 fl/- , Gpx4 fl/fl and Gpx4 GC−/− mice. (c) Appearance of Gpx4 fl/fl and Gpx4 GC−/− mice at 7 weeks. Representative images of three mice per group were shown. (d) Appearance of ovaries from Gpx4 fl/fl and Gpx4 GC−/− mice at 7 weeks. Representative images of three pairs of ovaries per group were shown. (e) Quantification of body weight (g) and ovary weight (mg) of Gpx4 fl/fl and Gpx4 GC−/− mice at 7 weeks. Data were presented as mean ± SEM, n = 6. ∗ P < 0.05, Student's t -test. (f) Quantification of malondialdehyde (MDA) levels in ovarian tissues. Data were presented as violin plots with data points ( n = 6). ∗ P < 0.05, one-way ANOVA. (g) Representative photomicrographs of ovarian sections. Serial sections were stained with hematoxylin-eosin (HE; left panels), Masson trichrome (middle panels), and TUNEL assay (right panels). Asterisks indicate corpora lutea; black arrows indicate preantral follicles; yellow arrows indicate collagen deposits; white arrows indicate TUNEL-positive cells. (h) Quantification of ( g ). Box-and-whisker plots with data points ( n = 6). ∗ P < 0.05, one-way ANOVA. (i) Western blot analysis of GPX4, 4-HNE, α-SMA, and Collagen I (Col1α) expression in ovarian tissues. GAPDH served as a loading control. Blots are representative of two samples per group. (j) Quantification of ( i ). Data were presented as mean ± SEM, n = 6. ∗ P < 0.05, one-way ANOVA.

Article Snippet: The following primary antibodies were used: GPX4 (ab125066, Abcam, UK), 4HNE (A26085, ABclonal, China), α-SMA (BS70000, Bioworld), Col1α (BS1530, Bioworld), DNMT1 (MA5-16169, Invitrogen, USA), DNMT3a (49768S, CST, USA), DNMT3b (48488S, CST, USA), MBD2 (ab188474, Abcam, UK), MeCP2 (A5694, ABclonal, China), MBD1 (29998-1-AP, Proteintech, USA), MBD3 (A8905, ABclonal, China), MBD4 (11270-1-AP, Proteintech, USA), FSHR (AF04761, AiFang Biological, China), Cyp19a1 (AF0449, AiFang Biological, China), MAZ (21068-1-AP, Proteintech, USA), NCoR (20018-1-AP, Proteintech, USA), HDAC3 (A19537, ABclonal, China), and GAPDH (FD0063, FDbio).

Techniques: Knock-Out, Agarose Gel Electrophoresis, Staining, TUNEL Assay, Whisker Assay, Western Blot, Expressing, Control

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: MBD2 inhibition by KCC-07 alleviates lipid peroxidation in granulosa cells. Primary granulosa cells (GCs) treated with DHEA (50 μM) in presence or absence with KCC-07 (KCC, 10 μM), or with Liproxstatin-1 (Lip-1, 200 nM), or with 5-Azacytidine (5-Aza/5Az, 10 μM, 24 h), respectively. (a) Representative micrographs of C11-BODIPY assay (upper three panels) and TUNEL staining (below panel). (b) Quantification of ( a ). Data were presented as Box-and-whisker plots with data points ( n = 4). ∗ P < 0.05, two-way ANOVA. (c) Western blot analysis of 4-HNE, GPX4, FSHR, and Cyp19a1 protein expression in GCs. GAPDH was as a loading control. (d) Quantification of ( c ). Data were presented as mean ± SEM, n = 3. ∗ P < 0.05, one-way ANOVA.

Article Snippet: The following primary antibodies were used: GPX4 (ab125066, Abcam, UK), 4HNE (A26085, ABclonal, China), α-SMA (BS70000, Bioworld), Col1α (BS1530, Bioworld), DNMT1 (MA5-16169, Invitrogen, USA), DNMT3a (49768S, CST, USA), DNMT3b (48488S, CST, USA), MBD2 (ab188474, Abcam, UK), MeCP2 (A5694, ABclonal, China), MBD1 (29998-1-AP, Proteintech, USA), MBD3 (A8905, ABclonal, China), MBD4 (11270-1-AP, Proteintech, USA), FSHR (AF04761, AiFang Biological, China), Cyp19a1 (AF0449, AiFang Biological, China), MAZ (21068-1-AP, Proteintech, USA), NCoR (20018-1-AP, Proteintech, USA), HDAC3 (A19537, ABclonal, China), and GAPDH (FD0063, FDbio).

Techniques: Inhibition, TUNEL Assay, Staining, Whisker Assay, Western Blot, Expressing, Control

Journal: Stem Cells Translational Medicine

Article Title: Extracellular vesicles derived from differentiated granulosa-like cells restore the ovarian function of rats with premature ovarian insufficiency

doi: 10.1093/stcltm/szaf081

Figure Lengend Snippet: Granulosa cell marker proteins in rUCMSC-derived granulosa-like cells. (A) rUCMSC and GLC cell morphology. (B) Real-time PCR was used to analyze the gene expression levels of the stem cell pluripotency marker Oct4; the specific precursor granulosa cell marker Foxl2; and the mature granulosa cell markers Amhr2 , Cyp19a1 , and Fshr during the differentiation of rUCMSCs into granulosa cells. The values are the means ± standard errors of the means (SEMs) ( n = 5). The control (Crl) cells were undifferentiated rUCMSCs (Day 0). Gapdh was used as an internal control for quantification. * P < 0.05; ** P < 0.01. (C) Immunofluorescence images of granulosa cell marker proteins and stem cell pluripotency marker proteins. Bar: 100 µm. (D) AMH and E2 hormone secretion level of differentiated and undifferentiated rUCMSC in vitro ( n = 3). Control (Crl) cells are undifferentiated rUCMSCs. * P < 0.05.

Article Snippet: The primary antibodies included FOXL2 (Cat. No. A16244; Rabbit; ABclonal), OCT4 (Cat. No. SC-5279; Mouse; Santa Cruz), AMHR2 (Cat. No. SC‐377413; Mouse; Santa Cruz), CYP19A1 (Cat. No. SC-374176; Mouse; Santa Cruz), FSHR (Cat. No. A1480; Rabbit; ABclonal), CD29-PE (Cat. No.102207; Mouse; BioLegend), CD90-FITC (Cat. No. 206105; Mouse; BioLegend), CD45-PE (Cat. No. 202207; Mouse; BioLegend), CD34 (Cat. No. SC-7324; Mouse; Santa Cruz), and CD44 (Cat. No. SC-7297; Mouse; Santa Cruz).

Techniques: Marker, Derivative Assay, Real-time Polymerase Chain Reaction, Gene Expression, Control, Immunofluorescence, In Vitro

Journal: Frontiers in Veterinary Science

Article Title: MNQ derivative D19 alleviates LPS-induced inflammation and oxidative stress in sheep follicular granulosa cells through the GPX4 -mediated ferroptosis

doi: 10.3389/fvets.2025.1621738

Figure Lengend Snippet: D19 restoring LPS-suppressed steroid hormone production in GCs. (A) ELISA measurement of A 4 , E 2 , and P 4 levels in GCs. (B,C) qRT-PCR and Western blotting were employed to detect the relative expression levels of mRNA and protein, respectively, for genes involved in steroid hormone synthesis ( HSD17B4 , CYP19A1 , 3β-HSD , CYP11A1 , and STAR ). Data from at least three independent experiments are presented as mean ± SEM. Significant differences ( p < 0.05) are denoted by different letters.

Article Snippet: CYP19A1 , bs-0114R , Bioss , Rabbit , 1:500.

Techniques: Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot, Expressing

Journal: Frontiers in Veterinary Science

Article Title: MNQ derivative D19 alleviates LPS-induced inflammation and oxidative stress in sheep follicular granulosa cells through the GPX4 -mediated ferroptosis

doi: 10.3389/fvets.2025.1621738

Figure Lengend Snippet: D19 preventing GPX4 deficiency-induced disruption of steroidogenesis in GCs. (A) Levels of E 2 and P 4 in different treatment groups were measured using ELISA. (B) The relative mRNA and protein expression levels of steroid hormone synthesis-related genes ( HSD17B4 , CYP19A1 , 3β-HSD , CYP11A1 , and STAR ) were detected by qRT-PCR and Western blotting, respectively. All experiments were performed in triplicate, and data are presented as mean ± SEM. Different letters indicate statistically significant differences ( p < 0.05).

Article Snippet: CYP19A1 , bs-0114R , Bioss , Rabbit , 1:500.

Techniques: Disruption, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Western Blot