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

Journal: Reproduction (Cambridge, England)

Article Title: Comparative analysis of PI3K-AKT and MEK-ERK1/2 signaling-driven molecular changes in granulosa cells

doi: 10.1530/REP-24-0317

Figure Lengend Snippet: Antibodies.

Article Snippet: Aromatase , SM2222P , Acris (Origene), USA , Mouse.

Techniques:

Journal: Reproduction (Cambridge, England)

Article Title: Comparative analysis of PI3K-AKT and MEK-ERK1/2 signaling-driven molecular changes in granulosa cells

doi: 10.1530/REP-24-0317

Figure Lengend Snippet: Regulation of steroidogenesis by AKT and ERK pathways. (A) and (B) Estradiol and progesterone levels in the culture media of cells treated with FSH and IGF1 compared to the untreated control ( n = 6). (C) and (D) Estradiol and progesterone levels in the conditioned media of cells cultured with independent or combined treatment of inhibitors compared to DMSO control ( n = 5). Data in (A) and (B) are presented as mean ± SEM and analyzed by unpaired t -test. Data in (C) and (D) are presented as mean ± SEM and analyzed by one-way ANOVA followed by Tukey’s post hoc test. (E), (F), (G), (H) and (I) Quantification of mRNA levels in qPCR for FOXL2 , FSHR , CYP19A1 , STAR and HSD3B genes ( n = 5). Data in (E), (F), (G), (H) and (I) are presented as box plots and analyzed using one-way ANOVA followed by Tukey’s post hoc test. (J) Protein expression of aromatase, STAR and ACTB genes in FSH- and IGF1-treated cells incubated with DMSO, PD98059 and LY294002 in three different replicates. (K) Graphical representation of the regulation of estradiol and progesterone production by ERK and AKT pathways in granulosa cells under the stimulation of FSH and IGF1. This graphic was generated using bioRender.com. Probability values <0.05 were considered statistically significant and are designated with up to four asterisk symbols to inform the strength of significant difference (* P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001). ns, not significant. The n value indicates the number of independent cell culture experiments analyzed using granulosa cells collected on different days.

Article Snippet: Aromatase , SM2222P , Acris (Origene), USA , Mouse.

Techniques: Control, Cell Culture, Expressing, Incubation, Generated

Journal: Biomaterials Research

Article Title: Injectable Biomimetic Hydrogel Constructs for Cell-Based Menopausal Hormone Therapy with Reduced Breast Cancer Potential

doi: 10.34133/bmr.0054

Figure Lengend Snippet: Characterization of mechanically isolated hormone-producing ovarian cells. (A) Hormone-producing ovarian cells are isolated by microdissection from the E 2 -primed ovary of 3-week-old female rats. (B) Morphology of granulosa cells (GCs) and theca cells (TCs) during a 72-h culture. Scale bars, 100 μm. (C) Fluorescence images of the staining of GC and TC with anti-CYP19 granulosa cell marker (green), anti-CYP17A1 theca cell marker (green), and DAPI (blue) nuclear counterstain. Scale bars, 20 μm. (D) The quantitative analysis of cell purity after staining of anti-CYP19 granulosa cell marker and anti-CYP17A1 theca cell marker, respectively, using flow cytometry.

Article Snippet: The primary antibodies used in this study are anti-CYP19 (BS-1292R, Bioss, Woburn, MA, USA) and anti-CYP17A1 (BS-54306R, Bioss).

Techniques: Isolation, Laser Capture Microdissection, Fluorescence, Staining, Marker, Flow Cytometry