quantitative pcr  (Bio-Rad)

Journal: Poultry Science

Article Title: Paeonol alleviates granulosa cell senescence in laying chickens via the PI3K/Akt/mTOR signaling pathway

doi: 10.1016/j.psj.2026.106750

Figure Lengend Snippet: Paeonol alleviates D-gal-induced apoptosis in GCs. A-E: qRT-PCR detection of BCL2 , Caspase-3, PCNA, CDK2 , and CCND1 . F-K: Western blot detection of Bax, Bcl-2, PCNA, Caspase-3, and CDK2 proteins. L-M: Annexin V/PI staining and flow cytometry analysis for detection of apoptosis; Q2: Annexin V⁺ / PI⁺, late apoptotic cells; Q3: Annexin V⁺ / PI⁻, early apoptotic cells. Data are presented as mean ± SEM ( n ≥ 3). Different lowercase letters indicate significant differences ( P < 0.05).

Article Snippet: Quantitative real-time PCR (qRT-PCR) was performed on a CFX96 Touch system (Bio-Rad, CA, USA) with 2 × SYBR qPCR Master Mix (Q711-02, Vazyme).

Techniques: Quantitative RT-PCR, Western Blot, Staining, Flow Cytometry

Journal: Poultry Science

Article Title: Paeonol alleviates granulosa cell senescence in laying chickens via the PI3K/Akt/mTOR signaling pathway

doi: 10.1016/j.psj.2026.106750

Figure Lengend Snippet: Paeonol alleviates D-gal-induced oxidative stress in GCs. A-B: Intracellular ROS was detected using DCFH-DA staining (green), Nuclei were counterstained with Hoechst 33342 (blue); Scale bar: 50 µm. C-G: qRT-PCR detection of antioxidant genes ( SOD, CAT, Mgst, Gsta , and Gsr ) . Data are presented as mean ± SEM ( n ≥ 3). Different lowercase letters indicate significant differences ( P < 0.05).

Article Snippet: Quantitative real-time PCR (qRT-PCR) was performed on a CFX96 Touch system (Bio-Rad, CA, USA) with 2 × SYBR qPCR Master Mix (Q711-02, Vazyme).

Techniques: Staining, Quantitative RT-PCR

Journal: Poultry Science

Article Title: Paeonol alleviates granulosa cell senescence in laying chickens via the PI3K/Akt/mTOR signaling pathway

doi: 10.1016/j.psj.2026.106750

Figure Lengend Snippet: Paeonol alleviates D-gal-induced mitochondrial dysfunction in GCs. A: Mitochondrial structure of GCs; Scale bars: 700 nm or 200 nm; Red arrow: mitochondria. B-C: Comparison of MDA and GSH levels in GCs. D-H: Western blot detection of the TOMM20, Drp1, MFN2 and OPA1. I-L: qRT-PCR analysis of mitochondrial gene expression-related genes ( TFAM, TFB2M, POLRMT and ATPase8 ). M-N: JC-1 staining for ΔΨm assessment; JC-1 aggregates (red) indicate high ΔΨm, whereas JC-1 monomers (green) indicate low ΔΨm; The red/green fluorescence ratio was quantified. Scale bar: 100 µm. O-P: Flow cytometry of ΔΨm and analysis of JC-1 aggregates/monomers ratio; Q2: JC-1 aggregates; Q3: JC-1 monomers. Data are presented as mean ± SEM ( n ≥ 3). Different lowercase letters indicate significant differences ( P < 0.05).

Article Snippet: Quantitative real-time PCR (qRT-PCR) was performed on a CFX96 Touch system (Bio-Rad, CA, USA) with 2 × SYBR qPCR Master Mix (Q711-02, Vazyme).

Techniques: Comparison, Western Blot, Quantitative RT-PCR, Gene Expression, Staining, Fluorescence, Flow Cytometry

Journal: Poultry Science

Article Title: Paeonol alleviates granulosa cell senescence in laying chickens via the PI3K/Akt/mTOR signaling pathway

doi: 10.1016/j.psj.2026.106750

Figure Lengend Snippet: Effects of Paeonol on follicular development, egg production and oxidative stress in late-laying chickens. A: Arrangement of ovaries and ovarian follicles (scale bar: 2 cm). B: The number of follicles; Pre: pre-ovulatory follicles; LYF: large yellow follicles; SYF: small yellow follicles; LWF: little white follicles; SWF: small white follicles; AF: atretic follicles. C: Cumulative egg production. D: E 2 levels in plasma. E-F: H&E staining (scale bar: 200 µm) and TUNEL staining (scale bar: 100 µm) of GCs; The percentage of TUNEL-positive cells (green) was calculated relative to total nuclei (DAPI); Red arrow: granulosa layer. G: Comparison of T-AOC levels in GCs. H-K: qRT-PCR detection of antioxidant enzyme genes ( SOD, CAT, Mgst , and Gsr) in GCs. Data are presented as mean ± SEM ( n ≥ 3). Different lowercase letters indicate significant differences ( P < 0.05).

Article Snippet: Quantitative real-time PCR (qRT-PCR) was performed on a CFX96 Touch system (Bio-Rad, CA, USA) with 2 × SYBR qPCR Master Mix (Q711-02, Vazyme).

Techniques: Clinical Proteomics, Staining, TUNEL Assay, Comparison, Quantitative RT-PCR

Journal: Poultry Science

Article Title: Paeonol alleviates granulosa cell senescence in laying chickens via the PI3K/Akt/mTOR signaling pathway

doi: 10.1016/j.psj.2026.106750

Figure Lengend Snippet: Effects of Paeonol on cell senescence and mitochondrial function in GCs of late-laying chickens. A-F: Western blot analysis of p16, TOMM20, Drp1, MFN2, and OPA1 protein expression in GCs of late-laying chickens. G-J: qRT-PCR analysis of mitochondrial gene expression-related genes ( TFAM, TFB2M, POLRMT and ATPase8 ) in GCs of late-laying chickens. Data are presented as mean ± SEM ( n ≥ 3). Different lowercase letters indicate significant differences ( P < 0.05).

Article Snippet: Quantitative real-time PCR (qRT-PCR) was performed on a CFX96 Touch system (Bio-Rad, CA, USA) with 2 × SYBR qPCR Master Mix (Q711-02, Vazyme).

Techniques: Western Blot, Expressing, Quantitative RT-PCR, Gene Expression