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rt cdna synthesis mix with oligo dt  (Solis BioDyne)


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    Structured Review

    Solis BioDyne rt cdna synthesis mix with oligo dt
    Rt Cdna Synthesis Mix With Oligo Dt, supplied by Solis BioDyne, used in various techniques. Bioz Stars score: 95/100, based on 51 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/rt+cdna+synthesis+mix/FIREScript+RT+cDNA+synthesis+MIX/pm41888742-71-11-10
    Average 95 stars, based on 51 article reviews
    rt cdna synthesis mix with oligo dt - by Bioz Stars, 2026-09
    95/100 stars

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    Related Articles

    Reverse Transcription:

    Article Title: RepE Monomer–Dimer Equilibrium Shapes Replication and Autoregulatory Control of the F Plasmid
    Article Snippet: Quantitative PCR reactions were performed with HOT FIREPol® EvaGreen® qPCR Supermix (Solis BioDyne, Tartu, Estonia) according to the manufacturer’s instructions. .. Reverse transcription and cDNA synthesis were performed with the FIREScript RT cDNA Synthesis Mix with Random Primers (Solis BioDyne, Tartu, Estonia) following the manufacturer’s protocol. ..

    Article Title: RepE Monomer–Dimer Equilibrium Shapes Replication and Autoregulatory Control of the F Plasmid
    Article Snippet: Quantitative PCR reactions were performed with HOT FIREPol ® EvaGreen ® qPCR Supermix (Solis BioDyne, Tartu, Estonia) according to the manufacturer’s instructions. .. Reverse transcription and cDNA synthesis were performed with the FIREScript RT cDNA Synthesis Mix with Random Primers (Solis BioDyne, Tartu, Estonia) following the manufacturer’s protocol. ..

    Article Title: Phoenixin regulates proliferation and UCP1 expression in rat brown primary preadipocytes
    Article Snippet: This is a PDF of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability.. This version will undergo additional copyediting, typesetting and review before it is published in its final form.. As such, this version is no longer the Accepted Manuscript, but it is not yet the definitive Version of Record; we are providing this early version to give early visibility of the article.

    cDNA Synthesis:

    Article Title: RepE Monomer–Dimer Equilibrium Shapes Replication and Autoregulatory Control of the F Plasmid
    Article Snippet: Quantitative PCR reactions were performed with HOT FIREPol® EvaGreen® qPCR Supermix (Solis BioDyne, Tartu, Estonia) according to the manufacturer’s instructions. .. Reverse transcription and cDNA synthesis were performed with the FIREScript RT cDNA Synthesis Mix with Random Primers (Solis BioDyne, Tartu, Estonia) following the manufacturer’s protocol. ..

    Article Title: RepE Monomer–Dimer Equilibrium Shapes Replication and Autoregulatory Control of the F Plasmid
    Article Snippet: Quantitative PCR reactions were performed with HOT FIREPol ® EvaGreen ® qPCR Supermix (Solis BioDyne, Tartu, Estonia) according to the manufacturer’s instructions. .. Reverse transcription and cDNA synthesis were performed with the FIREScript RT cDNA Synthesis Mix with Random Primers (Solis BioDyne, Tartu, Estonia) following the manufacturer’s protocol. ..

    Article Title: Gene expression kinetics in Sepsis After Cardiac Surgery (SACS): a multicentric prospective observational study
    Article Snippet: RNA was quantified using the NanoDropTM1000 spectrophotometer (NanoDrop Technologies, Inc). .. RNA was retro-transcribed in cDNA using FIREScript® RT cDNA synthesis MIX (Solis Biodyne) and used at a final concentration of 10 ng. .. For qPCR assay, Power SYBR Green PCR Master Mix (Applied Biosystems®) was used following the standard procedure indicated by Applied Biosystem®.

    Article Title: Soybean Cyst Nematode‐Resistant Protein AAT Rhg1 Affects Amino Acid Homeostasis and Betalain Accumulation
    Article Snippet: Frozen samples were homogenized with 2‐mm zirconium silicate grinding beads and PowerLyzer 24 (MO BIO Laboratories Inc., Carlsbad, CA, USA). .. Plant RNA was extracted using Direct‐zol RNA MiniPrep Plus Kits from Zymo Research (Irvine, CA, USA) following the manufacturer's instructions. cDNA was synthesized using FIREScript RT cDNA synthesis MIX from Solis BioDyne (Tartu, Estonia). .. Quantitative reverse transcription polymerase chain reactions (RT‐qPCR) were conducted to measure transcript abundance levels for RUBY and Rhg1‐GmAAT genes (Table ) using HOT FIREPol EvaGreen qPCR Supermix from Solis BioDyne and the CFX96 real‐time PCR detection system (BioRad, Hercules, CA, USA).

    Article Title: Crosstalk between TGF-β and Wnt/β-catenin signaling drives fibrogenic and stem-like phenotypes in senescent MDA-MB-231 breast cancer cells
    Article Snippet: RNA was extracted from frozen cell pellets using the Macherey-NagelTM NucleoSpinTM RNA Mini Kit, following the manufacturer’s instructions. .. RNA concentration and sample purity were measured using the NanoDrop ND-1000 spectrophotometer (Thermo Fisher). cDNA was synthesized from 250 to 500 ng of RNA using the FireScript RT cDNA Synthesis Mix (06-20-00100, Solis BioDyne) according to the manufacturer’s recommendations. ..

    Article Title: Phoenixin regulates proliferation and UCP1 expression in rat brown primary preadipocytes
    Article Snippet: This is a PDF of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability.. This version will undergo additional copyediting, typesetting and review before it is published in its final form.. As such, this version is no longer the Accepted Manuscript, but it is not yet the definitive Version of Record; we are providing this early version to give early visibility of the article.

    Article Title: AAT Rhg1 is a tonoplast protein that alters amino acid, metabolic and defense responses and nematode resistance
    Article Snippet: .. RNA from flash-frozen root samples was extracted using the Direct-zolTM RNA MiniPrep Plus Kit (Zymo Research, Irvine, CA, USA; see Supplemental Information). cDNA was synthesized using FIREScript ® RT cDNA synthesis MIX from Solis BioDyne (Tartu, Estonia). .. RNA from flash-frozen root samples was extracted using the Direct-zol RNA MiniPrep Plus Kit (Zymo Research, Irvine, CA, USA; see Supplemental Information).

    Concentration Assay:

    Article Title: Gene expression kinetics in Sepsis After Cardiac Surgery (SACS): a multicentric prospective observational study
    Article Snippet: RNA was quantified using the NanoDropTM1000 spectrophotometer (NanoDrop Technologies, Inc). .. RNA was retro-transcribed in cDNA using FIREScript® RT cDNA synthesis MIX (Solis Biodyne) and used at a final concentration of 10 ng. .. For qPCR assay, Power SYBR Green PCR Master Mix (Applied Biosystems®) was used following the standard procedure indicated by Applied Biosystem®.

    Article Title: Crosstalk between TGF-β and Wnt/β-catenin signaling drives fibrogenic and stem-like phenotypes in senescent MDA-MB-231 breast cancer cells
    Article Snippet: RNA was extracted from frozen cell pellets using the Macherey-NagelTM NucleoSpinTM RNA Mini Kit, following the manufacturer’s instructions. .. RNA concentration and sample purity were measured using the NanoDrop ND-1000 spectrophotometer (Thermo Fisher). cDNA was synthesized from 250 to 500 ng of RNA using the FireScript RT cDNA Synthesis Mix (06-20-00100, Solis BioDyne) according to the manufacturer’s recommendations. ..

    other:

    Article Title: Crosstalk between TGF-β and Wnt/β-catenin signaling drives fibrogenic and stem-like phenotypes in senescent MDA-MB-231 breast cancer cells.
    Article Snippet: RNA was extracted from frozen cell pellets using the Macherey-Nagel NucleoSpin RNA Mini Kit, following the manufacturer's instructions.

    Synthesized:

    Article Title: Soybean Cyst Nematode‐Resistant Protein AAT Rhg1 Affects Amino Acid Homeostasis and Betalain Accumulation
    Article Snippet: Frozen samples were homogenized with 2‐mm zirconium silicate grinding beads and PowerLyzer 24 (MO BIO Laboratories Inc., Carlsbad, CA, USA). .. Plant RNA was extracted using Direct‐zol RNA MiniPrep Plus Kits from Zymo Research (Irvine, CA, USA) following the manufacturer's instructions. cDNA was synthesized using FIREScript RT cDNA synthesis MIX from Solis BioDyne (Tartu, Estonia). .. Quantitative reverse transcription polymerase chain reactions (RT‐qPCR) were conducted to measure transcript abundance levels for RUBY and Rhg1‐GmAAT genes (Table ) using HOT FIREPol EvaGreen qPCR Supermix from Solis BioDyne and the CFX96 real‐time PCR detection system (BioRad, Hercules, CA, USA).

    Article Title: Crosstalk between TGF-β and Wnt/β-catenin signaling drives fibrogenic and stem-like phenotypes in senescent MDA-MB-231 breast cancer cells
    Article Snippet: RNA was extracted from frozen cell pellets using the Macherey-NagelTM NucleoSpinTM RNA Mini Kit, following the manufacturer’s instructions. .. RNA concentration and sample purity were measured using the NanoDrop ND-1000 spectrophotometer (Thermo Fisher). cDNA was synthesized from 250 to 500 ng of RNA using the FireScript RT cDNA Synthesis Mix (06-20-00100, Solis BioDyne) according to the manufacturer’s recommendations. ..

    Article Title: AAT Rhg1 is a tonoplast protein that alters amino acid, metabolic and defense responses and nematode resistance
    Article Snippet: .. RNA from flash-frozen root samples was extracted using the Direct-zolTM RNA MiniPrep Plus Kit (Zymo Research, Irvine, CA, USA; see Supplemental Information). cDNA was synthesized using FIREScript ® RT cDNA synthesis MIX from Solis BioDyne (Tartu, Estonia). .. RNA from flash-frozen root samples was extracted using the Direct-zol RNA MiniPrep Plus Kit (Zymo Research, Irvine, CA, USA; see Supplemental Information).

    Spectrophotometry:

    Article Title: Crosstalk between TGF-β and Wnt/β-catenin signaling drives fibrogenic and stem-like phenotypes in senescent MDA-MB-231 breast cancer cells
    Article Snippet: RNA was extracted from frozen cell pellets using the Macherey-NagelTM NucleoSpinTM RNA Mini Kit, following the manufacturer’s instructions. .. RNA concentration and sample purity were measured using the NanoDrop ND-1000 spectrophotometer (Thermo Fisher). cDNA was synthesized from 250 to 500 ng of RNA using the FireScript RT cDNA Synthesis Mix (06-20-00100, Solis BioDyne) according to the manufacturer’s recommendations. ..

    Real-time Polymerase Chain Reaction:

    Article Title: Phoenixin regulates proliferation and UCP1 expression in rat brown primary preadipocytes
    Article Snippet: This is a PDF of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability.. This version will undergo additional copyediting, typesetting and review before it is published in its final form.. As such, this version is no longer the Accepted Manuscript, but it is not yet the definitive Version of Record; we are providing this early version to give early visibility of the article.



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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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    Image Search Results


    Genetic analysis and expression of SURF1 gene variants in Case 2. (A) The probands 2 (P2) intronic variant c.515 + 3G > C caused abnormal splicing of the SURF1 gene, leading to the deletion of exon 5. (B) The expression level of the SURF1 gene in probands 2 (P2) was significantly lower than those in their parents. (C) PCR results of cDNA from the nuclear family of probands 2 (P2), the amplicon in the proband was shorter than the corresponding bands in the parents.

    Journal: Frontiers in Neurology

    Article Title: Study on Leigh syndrome caused by SURF1 gene mutations and its mechanisms

    doi: 10.3389/fneur.2026.1793054

    Figure Lengend Snippet: Genetic analysis and expression of SURF1 gene variants in Case 2. (A) The probands 2 (P2) intronic variant c.515 + 3G > C caused abnormal splicing of the SURF1 gene, leading to the deletion of exon 5. (B) The expression level of the SURF1 gene in probands 2 (P2) was significantly lower than those in their parents. (C) PCR results of cDNA from the nuclear family of probands 2 (P2), the amplicon in the proband was shorter than the corresponding bands in the parents.

    Article Snippet: Total RNA was extracted from patients and their parents using the Qiagen RNA Preparation Kit, and complementary DNA (cDNA) was synthesized by reverse transcription using the PrimeScriptTM Strand cDNA Synthesis Kit/RT Master Mix (TAKARA).

    Techniques: Expressing, Variant Assay, Amplification

    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