qrt pcr  (Roche)

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

Journal: PLOS One

Article Title: BaEV-pseudotyped lentiviral vectors enable stable CAR expression and cytotoxic function in NK cells

doi: 10.1371/journal.pone.0348674

Figure Lengend Snippet: (A) Schematic representation of the transfer vector used in this study. (B) Average virus titer (viral genome copies/mL) of each LV (n = 5). (C) A total of 2.5x10 5 NK92 cells were transduced with each type of lentiviral particle using 2.32x10 9 viral genome copies. The CAR expression level of each CAR-NK92 cell group was assessed by CD19 scFv-positive cell percentage on days 3, 7, and 14 post-transduction (day 3, n = 4; days 7 and 14, n = 5). (D) Cytotoxic activity of untransduced (UTD) and CAR-NK92 cells against K562 and Nalm-6. Cells were co-cultured for 4 hours at indicated E:T ratios (n = 4). (E) qRT-PCR analysis of the relative gene expression of CAR (n = 3). Data are shown as relative CD19 scFv gene expression using human GAPDH as a reference gene with analysis by the 2-ΔΔC T algorithm. (F) Gel electrophoresis analysis of CD19 scFv and human GAPDH amplified from genomic DNA of both UTD and CAR-NK92 cells at days 7 and 14 after transduction. DNA was extracted as total cellular genomic DNA, which may also contain non-integrated residual viral genomes. Data are presented as mean ± standard deviation (SD). Statistical analysis was performed using two-way ANOVA with Tukey’s multiple comparisons test (C), and one-way ANOVA with Tukey’s multiple comparisons test (B, E). *, p < 0.05; **, p < 0.01; ****, p < 0.0001; ns, non-significant.

Article Snippet: Subsequently, the number of viral copies was quantified using the Lenti-X qRT-PCR Titration Kit (631235, Takara Bio Inc.), and the titer of virus stock was measured by multiplying the quantified number by the fold dilution of the stock.

Techniques: Plasmid Preparation, Virus, Transduction, Expressing, Activity Assay, Cell Culture, Quantitative RT-PCR, Gene Expression, Nucleic Acid Electrophoresis, Amplification, Standard Deviation