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one step primescript iii rt qpcr mix  (TaKaRa)


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    Structured Review

    TaKaRa one step primescript iii rt qpcr mix
    One Step Primescript Iii Rt Qpcr Mix, supplied by TaKaRa, used in various techniques. Bioz Stars score: 96/100, based on 162 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/primescript+rt-qpcr+kit/One+Step+PrimeScript+III+RT-PCR+Kit/pm42087199-347-20-37
    Average 96 stars, based on 162 article reviews
    one step primescript iii rt qpcr mix - by Bioz Stars, 2026-10
    96/100 stars

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    Related Articles

    Quantitative RT-PCR:

    Article Title: SARS-CoV-2 infection is associated with hypothalamic orexin suppression and persistent cortical NeuN attenuation.
    Article Snippet: Total RNA was extracted from tissues using the Maxwell RSC Simply RNA Tissue Kit (AS1340; Promega, Madison, WI, USA) according to the manufacturer’s instructions. .. Quantitative RT-PCR was performed on a QuantStudio 3 Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) using either the One-Step PrimeScript III RT-qPCR Mix (RR600A) or the One Step TB Green PrimeScript RT-PCR Kit II (RR086A) (Takara, Kyoto, Japan), following the manufacturer’s protocols. .. Viral RNA targeting the SARS-CoV-2 nucleocapsid (N) gene was detected using the 2019-nCoV-N1 probe (10006770; Integrated DNA Technologies, Coralville, IA, USA).

    Article Title: Identification of Novel mRNA Biomarkers with Improved Performance for Colorectal Cancer Screening from a Multicenter Large Gene Screen
    Article Snippet: .. For RT-qPCR, Takara One Step PrimeScriptTM III RT-qPCR Mix with UNG (RR601A) was utilized for RT-qPCR. ..

    Article Title: Molecular epidemiology and genetic characteristics of enterovirus A identified in pediatric patients with hand, foot, and mouth disease and herpangina in Linyi, China, 2023-2024, following the relaxation of COVID-19 policies.
    Article Snippet: .. The RT-qPCR was performed in a 25 μL reaction volume using the One Step PrimeScriptTM III RT-qPCR Mix Kit (Code No. RR601A, Takara Bio ACCEPTED MANUSCRIPT AR TIC LE IN PR ES S ARTICLE IN PRESS Inc., Japan). ..

    Article Title: Molecular epidemiology and genetic characteristics of enterovirus A identified in pediatric patients with hand, foot, and mouth disease and herpangina in Linyi, China, 2023–2024, following the relaxation of COVID-19 policies
    Article Snippet: .. The RT-qPCR was performed in a 25 μL reaction volume using the One Step PrimeScriptTM III RT-qPCR Mix Kit (Code No. RR601A, Takara Bio Inc., Japan). ..

    Article Title: Limonin induces ferroptosis in cervical squamous cell carcinoma by activating the expression of soluble epoxide hydrolase 2 protein
    Article Snippet: The polymerase chain reaction (PCR) instrument (A37835; Thermo Fisher Scientific, China) was set at 42 °C for 5 min and 95 °C for 3 min to obtain the cDNA. .. A RT-qPCR kit (RR600A, TaKaRa Biotechnology Co., Ltd., China) was used to prepare the reaction system per the manufacturer’s instructions. .. The real-time fluorescence quantitative PCR instrument (A28139, Thermo Fisher Scientific, China) was set at 52 °C for 5 min, 95 °C for 10 s, followed by 40 cycles of 95 °C for 5 s and 63 °C for 30 s. The concentration time (CT) values were obtained after the reaction.

    Real-time Polymerase Chain Reaction:

    Article Title: SARS-CoV-2 infection is associated with hypothalamic orexin suppression and persistent cortical NeuN attenuation.
    Article Snippet: Total RNA was extracted from tissues using the Maxwell RSC Simply RNA Tissue Kit (AS1340; Promega, Madison, WI, USA) according to the manufacturer’s instructions. .. Quantitative RT-PCR was performed on a QuantStudio 3 Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) using either the One-Step PrimeScript III RT-qPCR Mix (RR600A) or the One Step TB Green PrimeScript RT-PCR Kit II (RR086A) (Takara, Kyoto, Japan), following the manufacturer’s protocols. .. Viral RNA targeting the SARS-CoV-2 nucleocapsid (N) gene was detected using the 2019-nCoV-N1 probe (10006770; Integrated DNA Technologies, Coralville, IA, USA).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: SARS-CoV-2 infection is associated with hypothalamic orexin suppression and persistent cortical NeuN attenuation.
    Article Snippet: Total RNA was extracted from tissues using the Maxwell RSC Simply RNA Tissue Kit (AS1340; Promega, Madison, WI, USA) according to the manufacturer’s instructions. .. Quantitative RT-PCR was performed on a QuantStudio 3 Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) using either the One-Step PrimeScript III RT-qPCR Mix (RR600A) or the One Step TB Green PrimeScript RT-PCR Kit II (RR086A) (Takara, Kyoto, Japan), following the manufacturer’s protocols. .. Viral RNA targeting the SARS-CoV-2 nucleocapsid (N) gene was detected using the 2019-nCoV-N1 probe (10006770; Integrated DNA Technologies, Coralville, IA, USA).

    Article Title: Detection of rabies virus RNA in dog-bite wounds in a rabies-endemic area: evidence from an observational cohort study.
    Article Snippet: .. A heminested broadly reactive lyssavirus PCR was used targeting the beginning of the Ngene (nucleotide position 55–641), 21 using the OneStep PrimeScript III RT-PCR kit (Takara Bio Inc., Shiga, Japan) and using in-house optimised cycles for the first PCR (30 min 50 ◦ C, followed by 40 cycles (2 min 94 ◦ C/ www.thelancet.com Vol ▪ ▪, 2026 3 30 sec 94 ◦ C, 30 sec 50 ◦ C, 30 sec 72 ◦ C, 605 bp product), 5 min 72 ◦ C) and Q5 HiFidelity DNA Polymerase (Sigma Aldrich, St.Louis, MO, USA) for the second, nested, PCR (40 cycles (30 sec 98 ◦ C/10 sec 98 ◦ C, 30 sec 50 ◦ C, 30 sec 72 ◦ C)/2 min 72 ◦ C, 581 bp product). .. A sequencing PCR (25 cycles of 1 min 96 ◦ C/10 sec 96 ◦ C, 5 sec 50 ◦ C, 1 min 60 ◦ C) was performed on 1–40 ng of PCR product using the primers from the second, nested, PCR, and the BigDye terminator V3.1 Cycle Sequencing Kit (Thermo Fisher Scientific, Waltham, MA, USA).

    SYBR Green Assay:

    Article Title: Combination of PARP and KRAS G12D inhibitors enhances therapeutic efficacy by exploiting vulnerabilities in PDAC
    Article Snippet: For cDNA synthesis, 200–1000 ng of total RNA samples were used for reverse transcription in 20 μl of reaction volume using the PrimeScript RT Reagent Kit (Takara, RR037). .. RT–qPCR was performed with the Power SYBR Green PCR Master Mix (Takara, RR601A). β-actin was used as a control for mRNA. ..

    Article Title: Combination of PARP and KRAS G12D inhibitors enhances therapeutic efficacy by exploiting vulnerabilities in PDAC.
    Article Snippet: For cDNA synthesis, 200-1,000 ng of total RNA samples were used for reverse transcription in 20ul of reaction volume using the PrimeScript RT Reagent Kit (Takara, RR037). .. RT–qPCR was performed with the Power SYBR Green PCR Master Mix (Takara, RR601A). ..

    Polymerase Chain Reaction:

    Article Title: Combination of PARP and KRAS G12D inhibitors enhances therapeutic efficacy by exploiting vulnerabilities in PDAC
    Article Snippet: For cDNA synthesis, 200–1000 ng of total RNA samples were used for reverse transcription in 20 μl of reaction volume using the PrimeScript RT Reagent Kit (Takara, RR037). .. RT–qPCR was performed with the Power SYBR Green PCR Master Mix (Takara, RR601A). β-actin was used as a control for mRNA. ..

    Article Title: Detection of rabies virus RNA in dog-bite wounds in a rabies-endemic area: evidence from an observational cohort study.
    Article Snippet: .. A heminested broadly reactive lyssavirus PCR was used targeting the beginning of the Ngene (nucleotide position 55–641), 21 using the OneStep PrimeScript III RT-PCR kit (Takara Bio Inc., Shiga, Japan) and using in-house optimised cycles for the first PCR (30 min 50 ◦ C, followed by 40 cycles (2 min 94 ◦ C/ www.thelancet.com Vol ▪ ▪, 2026 3 30 sec 94 ◦ C, 30 sec 50 ◦ C, 30 sec 72 ◦ C, 605 bp product), 5 min 72 ◦ C) and Q5 HiFidelity DNA Polymerase (Sigma Aldrich, St.Louis, MO, USA) for the second, nested, PCR (40 cycles (30 sec 98 ◦ C/10 sec 98 ◦ C, 30 sec 50 ◦ C, 30 sec 72 ◦ C)/2 min 72 ◦ C, 581 bp product). .. A sequencing PCR (25 cycles of 1 min 96 ◦ C/10 sec 96 ◦ C, 5 sec 50 ◦ C, 1 min 60 ◦ C) was performed on 1–40 ng of PCR product using the primers from the second, nested, PCR, and the BigDye terminator V3.1 Cycle Sequencing Kit (Thermo Fisher Scientific, Waltham, MA, USA).

    Article Title: Combination of PARP and KRAS G12D inhibitors enhances therapeutic efficacy by exploiting vulnerabilities in PDAC.
    Article Snippet: For cDNA synthesis, 200-1,000 ng of total RNA samples were used for reverse transcription in 20ul of reaction volume using the PrimeScript RT Reagent Kit (Takara, RR037). .. RT–qPCR was performed with the Power SYBR Green PCR Master Mix (Takara, RR601A). ..

    Control:

    Article Title: Combination of PARP and KRAS G12D inhibitors enhances therapeutic efficacy by exploiting vulnerabilities in PDAC
    Article Snippet: For cDNA synthesis, 200–1000 ng of total RNA samples were used for reverse transcription in 20 μl of reaction volume using the PrimeScript RT Reagent Kit (Takara, RR037). .. RT–qPCR was performed with the Power SYBR Green PCR Master Mix (Takara, RR601A). β-actin was used as a control for mRNA. ..

    Nested PCR:

    Article Title: Detection of rabies virus RNA in dog-bite wounds in a rabies-endemic area: evidence from an observational cohort study.
    Article Snippet: .. A heminested broadly reactive lyssavirus PCR was used targeting the beginning of the Ngene (nucleotide position 55–641), 21 using the OneStep PrimeScript III RT-PCR kit (Takara Bio Inc., Shiga, Japan) and using in-house optimised cycles for the first PCR (30 min 50 ◦ C, followed by 40 cycles (2 min 94 ◦ C/ www.thelancet.com Vol ▪ ▪, 2026 3 30 sec 94 ◦ C, 30 sec 50 ◦ C, 30 sec 72 ◦ C, 605 bp product), 5 min 72 ◦ C) and Q5 HiFidelity DNA Polymerase (Sigma Aldrich, St.Louis, MO, USA) for the second, nested, PCR (40 cycles (30 sec 98 ◦ C/10 sec 98 ◦ C, 30 sec 50 ◦ C, 30 sec 72 ◦ C)/2 min 72 ◦ C, 581 bp product). .. A sequencing PCR (25 cycles of 1 min 96 ◦ C/10 sec 96 ◦ C, 5 sec 50 ◦ C, 1 min 60 ◦ C) was performed on 1–40 ng of PCR product using the primers from the second, nested, PCR, and the BigDye terminator V3.1 Cycle Sequencing Kit (Thermo Fisher Scientific, Waltham, MA, USA).



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    PF effectively improves endothelial function (A) Hypoxia (1 % O 2 )-impaired HUVEC proliferation (CCK-8 assay) was rescued by PF (400 μM) at 48 h (n = 6). (B-D) Matrigel tube formation assay. (B) Representative images showing PF restored capillary-like structures under hypoxia. (C) Branching points and (D) total tube length quantified (n = 3). Scale bar: 100 μm. (E–K) PF modulated endothelial regulators in renal tissues. (E-G) eNOS, ANG-II and ET-1 mRNA by <t>RT-qPCR.</t> (H) Representative immunoblots and (I-K) quantification of eNOS, ANG-II and ET-1 protein (n = 5–6). (L–M) Serum levels of nitric oxide (NO, L) (n = 4) and ANG-II (M) (n = 6) detected by ELISA. Data presented as mean ± SEM. *p < 0.05, ** p < 0.01, *** p < 0.001 vs. CTL/Hypoxia; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. CRF/ Hypoxia + PF.
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    TaKaRa one step tb green primescript rt qpcr kit ii
    Validation of RNA-seq results <t>by</t> <t>RT-qPCR</t> in A549 cells transfected with miR-4430, miR-147a, or miR-1249-5p and infected with S. aureus USA300. (A) The Expression of STARD4 was significantly upregulated in cells treated with miR-147a and miR-1249-5p. (B) Expression of ITGA5 was significantly downregulated following treatment with miR-147a and miR-1249-5p. (C–E) Expression of PARP12 , STAT1 , and INSIG1 was significantly upregulated in cells treated with miR-4430. These results confirm the RNA-seq findings, supporting the downstream transcriptional effects of miR-4430 on immune signalling, whereas miR-147a and miR-1249-5p are associated with downstream modulation of adhesion- and lipid-related genes that influence S. aureus internalisation. Data represent three independent biological replicates (n = 3). * p < 0.05; ** p < 0.01; *** p < 0.001.
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    Image Search Results


    PF effectively improves endothelial function (A) Hypoxia (1 % O 2 )-impaired HUVEC proliferation (CCK-8 assay) was rescued by PF (400 μM) at 48 h (n = 6). (B-D) Matrigel tube formation assay. (B) Representative images showing PF restored capillary-like structures under hypoxia. (C) Branching points and (D) total tube length quantified (n = 3). Scale bar: 100 μm. (E–K) PF modulated endothelial regulators in renal tissues. (E-G) eNOS, ANG-II and ET-1 mRNA by RT-qPCR. (H) Representative immunoblots and (I-K) quantification of eNOS, ANG-II and ET-1 protein (n = 5–6). (L–M) Serum levels of nitric oxide (NO, L) (n = 4) and ANG-II (M) (n = 6) detected by ELISA. Data presented as mean ± SEM. *p < 0.05, ** p < 0.01, *** p < 0.001 vs. CTL/Hypoxia; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. CRF/ Hypoxia + PF.

    Journal: Journal of Advanced Research

    Article Title: Direct pharmacological targeting of Piezo1 by Paeoniflorin: a novel therapeutic approach for renal fibrosis

    doi: 10.1016/j.jare.2025.07.015

    Figure Lengend Snippet: PF effectively improves endothelial function (A) Hypoxia (1 % O 2 )-impaired HUVEC proliferation (CCK-8 assay) was rescued by PF (400 μM) at 48 h (n = 6). (B-D) Matrigel tube formation assay. (B) Representative images showing PF restored capillary-like structures under hypoxia. (C) Branching points and (D) total tube length quantified (n = 3). Scale bar: 100 μm. (E–K) PF modulated endothelial regulators in renal tissues. (E-G) eNOS, ANG-II and ET-1 mRNA by RT-qPCR. (H) Representative immunoblots and (I-K) quantification of eNOS, ANG-II and ET-1 protein (n = 5–6). (L–M) Serum levels of nitric oxide (NO, L) (n = 4) and ANG-II (M) (n = 6) detected by ELISA. Data presented as mean ± SEM. *p < 0.05, ** p < 0.01, *** p < 0.001 vs. CTL/Hypoxia; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. CRF/ Hypoxia + PF.

    Article Snippet: The RNA was then reverse-transcribed into cDNA using the ReverTra Ace qPCR RT Kit (6215A, TAKARA) according to the manufacturer's instructions.

    Techniques: CCK-8 Assay, Tube Formation Assay, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay

    PF attenuates Yoda1-induced Ca 2+ influx via functional modulation of Piezo1 channels (A) Immunofluorescence co-localization of Piezo1 (green) and PECAM-1 (red) in glomerular endothelium. Nuclei: DAPI (blue). Scale bar: 20 μm. (B-D) Renal Piezo1 expression analysis. (B) Piezo1 mRNA levels by RT-qPCR. (C) Representative Western blot bands. (D) Quantitative protein analysis normalized to GAPDH. (n = 5–6) (E–F) Intracellular Ca 2+ flux in HUVECs. Fluo-3 AM (5 μM)-labeled cells pre-treated with PF (400 μM, 1 h) or vehicle, then stimulated with Yoda1 (5 μM). PF significantly attenuated Yoda1-induced Ca2 + influx. Images captured via confocal microscopy (λex/λem: 488/530 nm). Scale bar: 20 μm. (G–H) Quantification of baseline (K) and Peak relative value (L) (n = 6). (I-J) Molecular docking of PF to Piezo1. (I) 3D structure showing hydrogen bonds (yellow dashes) with CYS977/PHE984/LYS988/LYS1154 and ARG1295. (J) 2D interaction map highlighting hydrophobic contacts and polar interactions. (K-L) Surface plasmon resonance (SPR) analysis. (K) Binding kinetics showing KD = 2.44 μM. (L) Sensorgrams of concentration-dependent binding.(M−Q) EndMT marker expression in HUVECs. (M) Western blot analysis of (N) eNOS, (O) VE-cadherin, (P) Vimentin, and (Q) TGF-β1. Quantification normalized to GAPDH (n = 3). Data presented as mean ± SEM. *p < 0.05, ** p < 0.01, *** p < 0.001 vs. CTL/Yoda1; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. CRF. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

    Journal: Journal of Advanced Research

    Article Title: Direct pharmacological targeting of Piezo1 by Paeoniflorin: a novel therapeutic approach for renal fibrosis

    doi: 10.1016/j.jare.2025.07.015

    Figure Lengend Snippet: PF attenuates Yoda1-induced Ca 2+ influx via functional modulation of Piezo1 channels (A) Immunofluorescence co-localization of Piezo1 (green) and PECAM-1 (red) in glomerular endothelium. Nuclei: DAPI (blue). Scale bar: 20 μm. (B-D) Renal Piezo1 expression analysis. (B) Piezo1 mRNA levels by RT-qPCR. (C) Representative Western blot bands. (D) Quantitative protein analysis normalized to GAPDH. (n = 5–6) (E–F) Intracellular Ca 2+ flux in HUVECs. Fluo-3 AM (5 μM)-labeled cells pre-treated with PF (400 μM, 1 h) or vehicle, then stimulated with Yoda1 (5 μM). PF significantly attenuated Yoda1-induced Ca2 + influx. Images captured via confocal microscopy (λex/λem: 488/530 nm). Scale bar: 20 μm. (G–H) Quantification of baseline (K) and Peak relative value (L) (n = 6). (I-J) Molecular docking of PF to Piezo1. (I) 3D structure showing hydrogen bonds (yellow dashes) with CYS977/PHE984/LYS988/LYS1154 and ARG1295. (J) 2D interaction map highlighting hydrophobic contacts and polar interactions. (K-L) Surface plasmon resonance (SPR) analysis. (K) Binding kinetics showing KD = 2.44 μM. (L) Sensorgrams of concentration-dependent binding.(M−Q) EndMT marker expression in HUVECs. (M) Western blot analysis of (N) eNOS, (O) VE-cadherin, (P) Vimentin, and (Q) TGF-β1. Quantification normalized to GAPDH (n = 3). Data presented as mean ± SEM. *p < 0.05, ** p < 0.01, *** p < 0.001 vs. CTL/Yoda1; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. CRF. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

    Article Snippet: The RNA was then reverse-transcribed into cDNA using the ReverTra Ace qPCR RT Kit (6215A, TAKARA) according to the manufacturer's instructions.

    Techniques: Functional Assay, Immunofluorescence, Expressing, Quantitative RT-PCR, Western Blot, Labeling, Confocal Microscopy, SPR Assay, Binding Assay, Concentration Assay, Marker

    PF inhibits matrix stiffness-induced acceleration of EndMT through Piezo1 activation (A–F) Matrix stiffness-dependent protein modulation in HUVECs. HUVECs were cultured on polyacrylamide hydrogels with soft (1.00 ± 0.31 kPa) or stiff (40.40 ± 2.39 kPa) for 48 h, with or without PF. (A) Western blot analysis of (B) Piezo1, (C) VE-Cadherin, (D) eNOS, (E) Vimentin, and (F) TGF-β1. Quantification normalized to GAPDH. (n = 3; *p < 0.05, **p < 0.01 vs. soft; #p < 0.05 vs. stiff without PF). (G-L) Piezo1 knockdown reverses stiffness-induced EndMT. HUVECs transfected with Piezo1 siRNA (50 nM, 24 h) or scramble siRNA (control) were cultured on stiffness hydrogels (40.40 ± 2.39 kPa) ± PF (400 μM). (G) Western blot analysis of (H) Piezo1, (I) VE-Cadherin, (J) eNOS, (K) Vimentin, and (L) TGF-β1. Quantification normalized to GAPDH (n = 3; *p < 0.05, **p < 0.01 vs. scramble siRNA control; ns vs. Piezo1 siRNA without PF).(M) Schematic of co-culture model. HUVECs and NRK-49F fibroblasts were co-cultured on stiffness-tunable hydrogels using a transwell system (0.4 μm pore size) for 5 days to assess paracrine signaling. (N) TGF-β1 levels in HUVEC supernatants from the co-culture system were quantified by ELISA. (n = 3; *p < 0.05 vs. softness; #p < 0.05 vs. stiffness without PF). (O) Fibrotic gene expression in NRK-49F cells. RT-qPCR analysis of Fibronectin , COL1A1 , Vimentin , and TGF-β1 mRNA levels. Data normalized to 18 s ( n = 3). (P-S) Fibrotic protein expression in NRK-49F cells. (P) Western blot analysis of (Q) Fibronectin, (R) COL1, (S) Vimentin, and (T) TGF-β1. Quantification normalized to GAPDH (n = 3). (U–V) Immunofluorescence of Fibronectin (red) in NRK-49F cells. Nuclei stained with DAPI (blue). (U) Representative images. (V) Quantification of fluorescence intensity using ImageJ (n = 3). Scale bar: 20 μm. Data presented as mean ± SEM. *p < 0.05, **p < 0.01 vs . softness co-culture; # p < 0.05 vs . stiffness co-culture without PF . (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

    Journal: Journal of Advanced Research

    Article Title: Direct pharmacological targeting of Piezo1 by Paeoniflorin: a novel therapeutic approach for renal fibrosis

    doi: 10.1016/j.jare.2025.07.015

    Figure Lengend Snippet: PF inhibits matrix stiffness-induced acceleration of EndMT through Piezo1 activation (A–F) Matrix stiffness-dependent protein modulation in HUVECs. HUVECs were cultured on polyacrylamide hydrogels with soft (1.00 ± 0.31 kPa) or stiff (40.40 ± 2.39 kPa) for 48 h, with or without PF. (A) Western blot analysis of (B) Piezo1, (C) VE-Cadherin, (D) eNOS, (E) Vimentin, and (F) TGF-β1. Quantification normalized to GAPDH. (n = 3; *p < 0.05, **p < 0.01 vs. soft; #p < 0.05 vs. stiff without PF). (G-L) Piezo1 knockdown reverses stiffness-induced EndMT. HUVECs transfected with Piezo1 siRNA (50 nM, 24 h) or scramble siRNA (control) were cultured on stiffness hydrogels (40.40 ± 2.39 kPa) ± PF (400 μM). (G) Western blot analysis of (H) Piezo1, (I) VE-Cadherin, (J) eNOS, (K) Vimentin, and (L) TGF-β1. Quantification normalized to GAPDH (n = 3; *p < 0.05, **p < 0.01 vs. scramble siRNA control; ns vs. Piezo1 siRNA without PF).(M) Schematic of co-culture model. HUVECs and NRK-49F fibroblasts were co-cultured on stiffness-tunable hydrogels using a transwell system (0.4 μm pore size) for 5 days to assess paracrine signaling. (N) TGF-β1 levels in HUVEC supernatants from the co-culture system were quantified by ELISA. (n = 3; *p < 0.05 vs. softness; #p < 0.05 vs. stiffness without PF). (O) Fibrotic gene expression in NRK-49F cells. RT-qPCR analysis of Fibronectin , COL1A1 , Vimentin , and TGF-β1 mRNA levels. Data normalized to 18 s ( n = 3). (P-S) Fibrotic protein expression in NRK-49F cells. (P) Western blot analysis of (Q) Fibronectin, (R) COL1, (S) Vimentin, and (T) TGF-β1. Quantification normalized to GAPDH (n = 3). (U–V) Immunofluorescence of Fibronectin (red) in NRK-49F cells. Nuclei stained with DAPI (blue). (U) Representative images. (V) Quantification of fluorescence intensity using ImageJ (n = 3). Scale bar: 20 μm. Data presented as mean ± SEM. *p < 0.05, **p < 0.01 vs . softness co-culture; # p < 0.05 vs . stiffness co-culture without PF . (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

    Article Snippet: The RNA was then reverse-transcribed into cDNA using the ReverTra Ace qPCR RT Kit (6215A, TAKARA) according to the manufacturer's instructions.

    Techniques: Activation Assay, Cell Culture, Western Blot, Knockdown, Transfection, Control, Co-Culture Assay, Pore Size, Enzyme-linked Immunosorbent Assay, Gene Expression, Quantitative RT-PCR, Expressing, Immunofluorescence, Staining, Fluorescence

    PF inhibits EndMT through the Piezo1-mediated HIF-1α signaling pathway (A–C) Renal HIF-1α expression analysis. (A) Representative RT-qPCR analysis of HIF-1α mRNA levels in kidney tissues. Data normalized to 18 s . (B-C) Western blot and quantification of HIF-1α protein expression in renal tissues. Data normalized to GAPDH (n = 5–6) . (D–H) Effects of PF or HIF-1α inhibitor BAY 87-2243 (10 μM, 24 h) on endothelial markers in HUVECs cultured with or without Piezo1 activation by Yoda1 (5 μM, 12 h). (D) Western blot analysis of (E) Piezo1, (F) HIF-1α, (G) VE-Cadherin, and (H) eNOS. Quantification normalized to GAPDH. Quantification showing Yoda1-induced Piezo1 upregulation and HIF-1α/VE-Cadherin/eNOS downregulation, reversed by PF or BAY 87-2243 (n = 3). (I-K) PF or BAY 87-2243 inhibits Yoda1-induced EndMT in HUVECs. (I) Western blot analysis of (J) Vimentin and (K) TGF-β1. Yoda1 increased Vimentin and TGF-β1, suppressed by PF or BAY 87-2243 (n = 3). (L) Schematic of HUVEC-NRK-49F co-culture. HUVECs pre-treated with/without Yoda1 (5 μM, 6 h) were co-cultured with NRK-49F fibroblasts for 48 h. (M) TGF-β1 levels in HUVEC supernatants from the co-culture system were quantified by ELISA. (N) RT-qPCR analysis of Fn1 , COL1A1 , and Vimentin mRNA in NRK-49F cells co-cultured with Yoda1-treated HUVECs. PF attenuated Yoda1-induced fibrotic marker expression (n = 3). (O-S) Western blot validation of (P) Fibronectin, (Q) COL1, (R) Vimentin, and (S) TGF-β1 in NRK-49F cells. PF reduced Yoda1-induced protein expression (n = 3). Data presented as mean ± SEM. *p < 0.05, ** p < 0.01, *** p < 0.001 vs. CTL/control; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. CRF/Yoda1.

    Journal: Journal of Advanced Research

    Article Title: Direct pharmacological targeting of Piezo1 by Paeoniflorin: a novel therapeutic approach for renal fibrosis

    doi: 10.1016/j.jare.2025.07.015

    Figure Lengend Snippet: PF inhibits EndMT through the Piezo1-mediated HIF-1α signaling pathway (A–C) Renal HIF-1α expression analysis. (A) Representative RT-qPCR analysis of HIF-1α mRNA levels in kidney tissues. Data normalized to 18 s . (B-C) Western blot and quantification of HIF-1α protein expression in renal tissues. Data normalized to GAPDH (n = 5–6) . (D–H) Effects of PF or HIF-1α inhibitor BAY 87-2243 (10 μM, 24 h) on endothelial markers in HUVECs cultured with or without Piezo1 activation by Yoda1 (5 μM, 12 h). (D) Western blot analysis of (E) Piezo1, (F) HIF-1α, (G) VE-Cadherin, and (H) eNOS. Quantification normalized to GAPDH. Quantification showing Yoda1-induced Piezo1 upregulation and HIF-1α/VE-Cadherin/eNOS downregulation, reversed by PF or BAY 87-2243 (n = 3). (I-K) PF or BAY 87-2243 inhibits Yoda1-induced EndMT in HUVECs. (I) Western blot analysis of (J) Vimentin and (K) TGF-β1. Yoda1 increased Vimentin and TGF-β1, suppressed by PF or BAY 87-2243 (n = 3). (L) Schematic of HUVEC-NRK-49F co-culture. HUVECs pre-treated with/without Yoda1 (5 μM, 6 h) were co-cultured with NRK-49F fibroblasts for 48 h. (M) TGF-β1 levels in HUVEC supernatants from the co-culture system were quantified by ELISA. (N) RT-qPCR analysis of Fn1 , COL1A1 , and Vimentin mRNA in NRK-49F cells co-cultured with Yoda1-treated HUVECs. PF attenuated Yoda1-induced fibrotic marker expression (n = 3). (O-S) Western blot validation of (P) Fibronectin, (Q) COL1, (R) Vimentin, and (S) TGF-β1 in NRK-49F cells. PF reduced Yoda1-induced protein expression (n = 3). Data presented as mean ± SEM. *p < 0.05, ** p < 0.01, *** p < 0.001 vs. CTL/control; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. CRF/Yoda1.

    Article Snippet: The RNA was then reverse-transcribed into cDNA using the ReverTra Ace qPCR RT Kit (6215A, TAKARA) according to the manufacturer's instructions.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Cell Culture, Activation Assay, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Marker, Biomarker Discovery, Control

    Validation of RNA-seq results by RT-qPCR in A549 cells transfected with miR-4430, miR-147a, or miR-1249-5p and infected with S. aureus USA300. (A) The Expression of STARD4 was significantly upregulated in cells treated with miR-147a and miR-1249-5p. (B) Expression of ITGA5 was significantly downregulated following treatment with miR-147a and miR-1249-5p. (C–E) Expression of PARP12 , STAT1 , and INSIG1 was significantly upregulated in cells treated with miR-4430. These results confirm the RNA-seq findings, supporting the downstream transcriptional effects of miR-4430 on immune signalling, whereas miR-147a and miR-1249-5p are associated with downstream modulation of adhesion- and lipid-related genes that influence S. aureus internalisation. Data represent three independent biological replicates (n = 3). * p < 0.05; ** p < 0.01; *** p < 0.001.

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Host-directed microRNA-based intervention against intracellular Staphylococcus aureus : high-throughput screening identifies miR-4430, miR-147a, and miR-1249-5p as multifunctional antimicrobial candidates

    doi: 10.3389/fcimb.2026.1772100

    Figure Lengend Snippet: Validation of RNA-seq results by RT-qPCR in A549 cells transfected with miR-4430, miR-147a, or miR-1249-5p and infected with S. aureus USA300. (A) The Expression of STARD4 was significantly upregulated in cells treated with miR-147a and miR-1249-5p. (B) Expression of ITGA5 was significantly downregulated following treatment with miR-147a and miR-1249-5p. (C–E) Expression of PARP12 , STAT1 , and INSIG1 was significantly upregulated in cells treated with miR-4430. These results confirm the RNA-seq findings, supporting the downstream transcriptional effects of miR-4430 on immune signalling, whereas miR-147a and miR-1249-5p are associated with downstream modulation of adhesion- and lipid-related genes that influence S. aureus internalisation. Data represent three independent biological replicates (n = 3). * p < 0.05; ** p < 0.01; *** p < 0.001.

    Article Snippet: Reverse transcription qPCR (RT-qPCR) was performed using the One-Step TB Green PrimeScript RT-qPCR Kit II (Takara Bio, Spain) on a QuantStudio 5 system (Applied Biosystems, USA).

    Techniques: Biomarker Discovery, RNA Sequencing, Quantitative RT-PCR, Transfection, Infection, Expressing