Review



p16 ink4a antibody f 12  (Santa Cruz Biotechnology)


Bioz Verified Symbol Santa Cruz Biotechnology is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 96
      Buy from Supplier

    Structured Review

    Santa Cruz Biotechnology p16 ink4a antibody f 12
    M6A modifications within total mRNA were significantly enhanced during the aging process . ( A ) Left: representative photographs of 2-month and 20-month-old male C57BL/6 mouse; Right: representative photographs of SA-β-gal staining in kidney tissues from young and old C57-BL/6 mice. ( B ) Left: representative photographs of SA-β-gal staining of P3 and P9 MEF cells; Right: the <t>p16</t> expression levels in P3 and P9 MEF cells. ( C ) Left: representative photographs of SA-β-gal staining of H 2 O 2 -induced premature senescence of NIH/3T3 cells; Right: the p16 expression levels in H 2 O 2 treated NIH/3T3 cells. ( D ) Poly(A)+ RNA was extracted from the brain and liver tissue of young and old C57-BL/6 mice and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( E ) Poly(A)+ RNA was extracted from the P3 and P9 MEF cells and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( F ) Poly(A)+ RNA was extracted from the P3 and P9 MEF cells and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( G ) Expression levels of m6A modification associated proteins were measured in P3 and P9 MEF cells, and the quantitative results were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; One sample t test (MEF P9 vs 1, METTL3: p =0.0027, t = 9.176).
    P16 Ink4a Antibody F 12, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 2196 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/p16+ink4a/p16+Antibody/pmc12834426-37-42-47
    Average 96 stars, based on 2196 article reviews
    p16 ink4a antibody f 12 - by Bioz Stars, 2026-10
    96/100 stars

    Images

    1) Product Images from "The Flip Side of the Coin: METTL3 Serves as a Novel Cellular Senescence Accelerator via Negative Regulation of ITGA9"

    Article Title: The Flip Side of the Coin: METTL3 Serves as a Novel Cellular Senescence Accelerator via Negative Regulation of ITGA9

    Journal: Aging and Disease

    doi: 10.14336/AD.2024.1715

    M6A modifications within total mRNA were significantly enhanced during the aging process . ( A ) Left: representative photographs of 2-month and 20-month-old male C57BL/6 mouse; Right: representative photographs of SA-β-gal staining in kidney tissues from young and old C57-BL/6 mice. ( B ) Left: representative photographs of SA-β-gal staining of P3 and P9 MEF cells; Right: the p16 expression levels in P3 and P9 MEF cells. ( C ) Left: representative photographs of SA-β-gal staining of H 2 O 2 -induced premature senescence of NIH/3T3 cells; Right: the p16 expression levels in H 2 O 2 treated NIH/3T3 cells. ( D ) Poly(A)+ RNA was extracted from the brain and liver tissue of young and old C57-BL/6 mice and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( E ) Poly(A)+ RNA was extracted from the P3 and P9 MEF cells and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( F ) Poly(A)+ RNA was extracted from the P3 and P9 MEF cells and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( G ) Expression levels of m6A modification associated proteins were measured in P3 and P9 MEF cells, and the quantitative results were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; One sample t test (MEF P9 vs 1, METTL3: p =0.0027, t = 9.176).
    Figure Legend Snippet: M6A modifications within total mRNA were significantly enhanced during the aging process . ( A ) Left: representative photographs of 2-month and 20-month-old male C57BL/6 mouse; Right: representative photographs of SA-β-gal staining in kidney tissues from young and old C57-BL/6 mice. ( B ) Left: representative photographs of SA-β-gal staining of P3 and P9 MEF cells; Right: the p16 expression levels in P3 and P9 MEF cells. ( C ) Left: representative photographs of SA-β-gal staining of H 2 O 2 -induced premature senescence of NIH/3T3 cells; Right: the p16 expression levels in H 2 O 2 treated NIH/3T3 cells. ( D ) Poly(A)+ RNA was extracted from the brain and liver tissue of young and old C57-BL/6 mice and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( E ) Poly(A)+ RNA was extracted from the P3 and P9 MEF cells and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( F ) Poly(A)+ RNA was extracted from the P3 and P9 MEF cells and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( G ) Expression levels of m6A modification associated proteins were measured in P3 and P9 MEF cells, and the quantitative results were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; One sample t test (MEF P9 vs 1, METTL3: p =0.0027, t = 9.176).

    Techniques Used: Staining, Expressing, Dot Blot, Control, Modification

    METTL3 accelerated replicative senescence in MEFs . ( A ) The protein levels in MEFs transfected with siNC and siMETTL3-1/2/3. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (siMETTL3-1 vs 1: p =0.0113, t =9.328; siMETTL3-2VS 1: p =0.0022, t =21.05; siMETTL3-3 vs 1: p =0.0117, t = 9.155). ( B ) Poly(A)+ RNA was extracted from MEF cells transfected with siNC and siMETTL3 and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( C ) Western blot assay of p16 expression in MEFs transfected with siNC and siMETTL3. The quantitative results were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; One sample t test (siMETTL3 vs 1, p =0.0006, t =15.36). ( D ) Representative photographs of SA-β-gal staining of MEF cells transfected with siNC and siMETTL3 (×100). The quantitative results of SA-β-gal staining were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; Paired t test (siMETTL3 vs siNC, p =0.0025, t =9.407). ( E ) Representative photographs of cells stained with DAPI (blue fluorescence) and BrdU (green fluorescence) in siNC and siMETTL3 transfected MEF cells (scale bar: 400μm). The quantitative results of the ratio (BrdU stained cells / DAPI stained cells) were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; Paired t test (siMETTL3 vs siNC, p =0.009, t =6.065). ( F ) The mRNA levels in MEFs infected with Lenti-NC and METTL3 lentivirus. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (METTL3VS 1, p =0.0058, t =13.03). ( G ) Poly(A)+ RNA was extracted from MEF cells infected with Lenti-NC and METTL3 lentivirus and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( H ) Western blot assay of METTL3 and p16 expression in MEFs infected with METTL3 lentivirus. The quantitative results were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (METTL3 vs 1, p =0.0342, t =5.271). ( I ) Representative photographs of SA-β-gal staining of MEF cells infected with Lenti-NC and METTL3 lentivirus (×100). The quantitative results of SA-β-gal staining were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test (METTL3 vs Lenti-NC, p =0.0088, t =10.59). ( J ) Representative photographs of cells stained with DAPI (blue fluorescence) and BrdU (green fluorescence) in Lenti-NC and METTL3 lentivirus infected MEF cells (scale bar: 400μm). The quantitative results of the ratio (BrdU stained cells / DAPI stained cells) were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test (METTL3 vs Lenti-NC, p =0.0065, t =12.35).
    Figure Legend Snippet: METTL3 accelerated replicative senescence in MEFs . ( A ) The protein levels in MEFs transfected with siNC and siMETTL3-1/2/3. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (siMETTL3-1 vs 1: p =0.0113, t =9.328; siMETTL3-2VS 1: p =0.0022, t =21.05; siMETTL3-3 vs 1: p =0.0117, t = 9.155). ( B ) Poly(A)+ RNA was extracted from MEF cells transfected with siNC and siMETTL3 and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( C ) Western blot assay of p16 expression in MEFs transfected with siNC and siMETTL3. The quantitative results were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; One sample t test (siMETTL3 vs 1, p =0.0006, t =15.36). ( D ) Representative photographs of SA-β-gal staining of MEF cells transfected with siNC and siMETTL3 (×100). The quantitative results of SA-β-gal staining were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; Paired t test (siMETTL3 vs siNC, p =0.0025, t =9.407). ( E ) Representative photographs of cells stained with DAPI (blue fluorescence) and BrdU (green fluorescence) in siNC and siMETTL3 transfected MEF cells (scale bar: 400μm). The quantitative results of the ratio (BrdU stained cells / DAPI stained cells) were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; Paired t test (siMETTL3 vs siNC, p =0.009, t =6.065). ( F ) The mRNA levels in MEFs infected with Lenti-NC and METTL3 lentivirus. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (METTL3VS 1, p =0.0058, t =13.03). ( G ) Poly(A)+ RNA was extracted from MEF cells infected with Lenti-NC and METTL3 lentivirus and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( H ) Western blot assay of METTL3 and p16 expression in MEFs infected with METTL3 lentivirus. The quantitative results were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (METTL3 vs 1, p =0.0342, t =5.271). ( I ) Representative photographs of SA-β-gal staining of MEF cells infected with Lenti-NC and METTL3 lentivirus (×100). The quantitative results of SA-β-gal staining were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test (METTL3 vs Lenti-NC, p =0.0088, t =10.59). ( J ) Representative photographs of cells stained with DAPI (blue fluorescence) and BrdU (green fluorescence) in Lenti-NC and METTL3 lentivirus infected MEF cells (scale bar: 400μm). The quantitative results of the ratio (BrdU stained cells / DAPI stained cells) were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test (METTL3 vs Lenti-NC, p =0.0065, t =12.35).

    Techniques Used: Transfection, Dot Blot, Staining, Control, Western Blot, Expressing, Fluorescence, Infection

    ITGA9 staved off senescence in MEFs . ( A ) The mRNA and protein levels of ITGA9 in MEF cells transfected with siNC and siITGA9-1/2/3.Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (siITGA9-1 vs 11: p =0.0002, t =68.12; siITGA9-2 vs 1: p =0.0003, t =63.15 ; siITGA9-3 vs 11: p =0.0111, t =9.410; ) (B) The p16 expression levels in MEF cells transfected with siNC and siITGA9. The quantitative results were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (siITGA9 vs 1, p =0.0012, t =28.41). ( C ) Representative photographs of SA-β-gal staining of MEF cells transfected with siNC and siITGA9 (×100). The quantitative results of SA-β-gal staining were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test (siITGA9 vs siNC, p =0.0026, t =19.65). ( D ) Representative photographs of cells stained with DAPI (blue fluorescence) and BrdU (green fluorescence) in siNC and siITGA9 transfected MEF cells (scale bar: 400μm). The quantitative results of the ratio ( BrdU stained cells / DAPI stained cells) were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test (siITGA9 vs siNC, p =0.0144, t =4.138). ( E ) Representative photographs of cells stained with DAPI (blue fluorescence) and BrdU (green fluorescence) in Lenti-NC and ITGA9 infected MEF cells (scale bar: 400μm). The quantitative results of the ratio ( BrdU stained cells / DAPI stained cells) were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; Paired t test (ITGA9 vs Lenti-NC, p =0.01, t =5.843). ( F ) The mRNA levels of ITGA9 in MEF cells infected with Lenti-NC or ITGA9 lentivirus. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (ITGA9 vs 1, p =0.0025, t =20.08). ( G ) The ITGA9 and p16 protein expression levels in MEFs infected with Lenti-NC and ITGA9 lentivirus. The results of grayscale scanning were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (ITGA9 vs 1, p =0.0256, t =6.124). ( H ) Representative photographs of SA-β-gal staining of MEF cells infected with Lenti-NC and ITGA9 lentivirus (×100). The quantitative results of SA-β-gal staining were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test (ITGA9 vs Lenti-NC, p =0.0009, t =33.30).
    Figure Legend Snippet: ITGA9 staved off senescence in MEFs . ( A ) The mRNA and protein levels of ITGA9 in MEF cells transfected with siNC and siITGA9-1/2/3.Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (siITGA9-1 vs 11: p =0.0002, t =68.12; siITGA9-2 vs 1: p =0.0003, t =63.15 ; siITGA9-3 vs 11: p =0.0111, t =9.410; ) (B) The p16 expression levels in MEF cells transfected with siNC and siITGA9. The quantitative results were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (siITGA9 vs 1, p =0.0012, t =28.41). ( C ) Representative photographs of SA-β-gal staining of MEF cells transfected with siNC and siITGA9 (×100). The quantitative results of SA-β-gal staining were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test (siITGA9 vs siNC, p =0.0026, t =19.65). ( D ) Representative photographs of cells stained with DAPI (blue fluorescence) and BrdU (green fluorescence) in siNC and siITGA9 transfected MEF cells (scale bar: 400μm). The quantitative results of the ratio ( BrdU stained cells / DAPI stained cells) were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test (siITGA9 vs siNC, p =0.0144, t =4.138). ( E ) Representative photographs of cells stained with DAPI (blue fluorescence) and BrdU (green fluorescence) in Lenti-NC and ITGA9 infected MEF cells (scale bar: 400μm). The quantitative results of the ratio ( BrdU stained cells / DAPI stained cells) were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; Paired t test (ITGA9 vs Lenti-NC, p =0.01, t =5.843). ( F ) The mRNA levels of ITGA9 in MEF cells infected with Lenti-NC or ITGA9 lentivirus. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (ITGA9 vs 1, p =0.0025, t =20.08). ( G ) The ITGA9 and p16 protein expression levels in MEFs infected with Lenti-NC and ITGA9 lentivirus. The results of grayscale scanning were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (ITGA9 vs 1, p =0.0256, t =6.124). ( H ) Representative photographs of SA-β-gal staining of MEF cells infected with Lenti-NC and ITGA9 lentivirus (×100). The quantitative results of SA-β-gal staining were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test (ITGA9 vs Lenti-NC, p =0.0009, t =33.30).

    Techniques Used: Transfection, Expressing, Staining, Fluorescence, Infection

    METTL3 accelerated cellular senescence by modulating ITGA9 . ( A ) The p16 level in MEF cells transfected with siNC, siMETTL3 or siMETTL3 and siITGA9. The quantitative results were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; Paired t test ( p =0.0058, t =7.062). ( B ) The p16 level in MEF cells infected with Lenti-NC, METTL3 or METTL3 and ITGA9. The quantitative results were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test ( p =0.017, t =7.563). (C, D) Representative photographs of SA-β-gal staining (C) and BrdU incorporation staining (D) of MEF cells transfected with siNC, siMETTL3, siITGA9, or siMETTL3 and siITGA9. The quantitative results were shown on the right. Mean (±SEM), passed normality test: Shapiro-Wilk test; Paired t test (C), n=3, p =0.0044, t =15.02; (D), n=4, p =0.0003, t =19.14). (E, F) Representative photographs of SA-β-gal staining (E) and BrdU incorporation staining (F) of MEF cells infected with Lenti-NC, METTL3, ITGA9 or METTL3 and ITGA9. The quantitative results were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test ((E): p =0.0095, t =10.18; (F): p =0.0231, t =6.469).
    Figure Legend Snippet: METTL3 accelerated cellular senescence by modulating ITGA9 . ( A ) The p16 level in MEF cells transfected with siNC, siMETTL3 or siMETTL3 and siITGA9. The quantitative results were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; Paired t test ( p =0.0058, t =7.062). ( B ) The p16 level in MEF cells infected with Lenti-NC, METTL3 or METTL3 and ITGA9. The quantitative results were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test ( p =0.017, t =7.563). (C, D) Representative photographs of SA-β-gal staining (C) and BrdU incorporation staining (D) of MEF cells transfected with siNC, siMETTL3, siITGA9, or siMETTL3 and siITGA9. The quantitative results were shown on the right. Mean (±SEM), passed normality test: Shapiro-Wilk test; Paired t test (C), n=3, p =0.0044, t =15.02; (D), n=4, p =0.0003, t =19.14). (E, F) Representative photographs of SA-β-gal staining (E) and BrdU incorporation staining (F) of MEF cells infected with Lenti-NC, METTL3, ITGA9 or METTL3 and ITGA9. The quantitative results were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test ((E): p =0.0095, t =10.18; (F): p =0.0231, t =6.469).

    Techniques Used: Transfection, Infection, Staining, BrdU Incorporation Assay

    Related Articles

    Staining:

    Article Title: β-catenin initiates peritoneal fibrosis by triggering mitochondrial fission-mediated mesothelial cell senescence fate transition
    Article Snippet: .. Sirius red staining was performed using the picro-red staining kit (PH1098; Scientific Phygene, China), at room temperature for 2 h. Paraffin Sects. (4 μm thickness) were performed by immunohistochemical staining with antibodies against fibronectin (FN; F3648; Sigma, USA; 1:200), p16 INK4A (sc-1661; Santa Cruz Biotechnology, USA; 1:100), Drp1 (ab184247; Abcam, USA; 1:50), and S100A4/FSP1 (ab218512; Abcam, USA; 1:50). ..

    Immunohistochemical staining:

    Article Title: β-catenin initiates peritoneal fibrosis by triggering mitochondrial fission-mediated mesothelial cell senescence fate transition
    Article Snippet: .. Sirius red staining was performed using the picro-red staining kit (PH1098; Scientific Phygene, China), at room temperature for 2 h. Paraffin Sects. (4 μm thickness) were performed by immunohistochemical staining with antibodies against fibronectin (FN; F3648; Sigma, USA; 1:200), p16 INK4A (sc-1661; Santa Cruz Biotechnology, USA; 1:100), Drp1 (ab184247; Abcam, USA; 1:50), and S100A4/FSP1 (ab218512; Abcam, USA; 1:50). ..

    Flow Cytometry:

    Article Title: Selective Targeting of Senescent FHs74Int Cells by Human Breast Milk Free Fatty Acids
    Article Snippet: The presence of senescence in cultured cells was assessed using Cayman’s Senescence-Associated β-Galactosidase staining kit according to the manufacturer’s instructions (Cayman Chemical Company, Ann Arbor, MI, USA). .. Flow cytometry analysis of p16 INK4a and p21 was performed using unconjugated anti-p16 INK4a and unconjugated p21 Waf1/Cip1 (F-5), both from Santa Cruz Biotechnology (Dallas, TX, USA). .. FITC-conjugated anti-mouse IgG (Thermo Fisher) was used as a secondary antibody, and the data were acquired using the BD Accuri C6 (BD Biosciences, San Jose, CA, USA).

    Article Title: Selective Targeting of Senescent FHs74Int Cells by Human Breast Milk Free Fatty Acids
    Article Snippet: The presence of senescence in cultured cells was assessed using Cayman’s SenescenceAssociated β-Galactosidase staining kit according to the manufacturer’s instructions (Cayman Chemical Company, Ann Arbor, MI, USA). .. Flow cytometry analysis of p16 INK4a and p21 was performed using unconjugated anti-p16 INK4a and unconjugated p21 Waf1/Cip1 (F-5), both from Santa Cruz Biotechnology (Dallas, TX, USA). .. FITC-conjugated anti-mouse IgG (Thermo Fisher) was used as a secondary antibody, and the data were acquired using the BD Accuri C6 (BD Biosciences, San Jose, CA, USA).

    Cell-Signaling:

    Article Title: Transient Receptor Potential Vanilloid 4 Knockdown Decreases Extracellular Matrix Synthesis via Autophagy Suppression in the Rat Intervertebral Disc
    Article Snippet: .. TRPV1 (rabbit, polyclonal) ACC-030 Almone labs (Jerusalem, Israel) TRPV4 (rabbit, polyclonal) ACC-124 Almone labs (Jerusalem, Israel) TRPV5 (rabbit, polyclonal) ACC-035 Almone labs (Jerusalem, Israel) AMPK (rabbit, polyclonal) 2532S Cell Signaling Technology (Danvers, MA) P-AMPK (rabbit, monoclonal) 2535S Cell Signaling Technology (Danvers, MA) mTOR (rabbit, monoclonal) 2983 Cell Signaling Technology (Danvers, MA) RAPTOR (rabbit, monoclonal) 2280 Cell Signaling Technology (Danvers, MA) RICTOR (rabbit, monoclonal) 2114 Cell Signaling Technology (Danvers, MA) p70/S6K (rabbit, monoclonal) 2708 Cell Signaling Technology (Danvers, MA) LC3 (rabbit, monoclonal) 3868 Cell Signaling Technology (Danvers, MA) p62/SQSTM1 (mouse, monoclonal) ab56416 Abcam (Cambridge, UK) Cleaved caspase-9 (rabbit, polyclonal) 9507 Cell Signaling Technology (Danvers, MA) PARP (rabbit, monoclonal) 9532 Cell Signaling Technology (Danvers, MA) Cleaved PARP (rabbit, monoclonal) 5625 Cell Signaling Technology (Danvers, MA) p53 (mouse, monoclonal) 2524 Cell Signaling Technology (Danvers, MA) p21/CIP1 (mouse, monoclonal) sc-6246 Santa Cruz Biotechnology (Santa Cruz, CA) Brachyury (mouse, monoclonal) sc-166962 Santa Cruz Biotechnology (Santa Cruz, CA) CD24 (mouse, monoclonal) sc-19585 Santa Cruz Biotechnology (Santa Cruz, CA) p16/INK4a (mouse, monoclonal) sc-1661 Santa Cruz Biotechnology (Santa Cruz, CA) COL2A1 (mouse, monoclonal) sc-52658 Santa Cruz Biotechnology (Santa Cruz, CA) Aggrecan (mouse, monoclonal) ab3778 Abcam (Cambridge, UK) MMP3 (rabbit, monoclonal) ab52915 Abcam (Cambridge, UK) MMP13 (rabbit, polyclonal) ab39012 Abcam (Cambridge, UK) TIMP1 (mouse, monoclonal) sc-21734 Santa Cruz Biotechnology (Santa Cruz, CA) TIMP2 (rabbit, monoclonal) 5738 Cell Signaling Technology (Danvers, MA) Tubulin (mouse, monoclonal) T9026 Sigma-Aldrich (St. Louis, MO) RNAi Lipofectamine RNAiMAX transfection reagent 13778150 Thermo Fisher Scientific (Waltham, MA) Invivofectamine 3.0 Reagent IVF3001 Thermo Fisher Scientific (Waltham, MA) Opti-minimal essential medium I 3198070 Thermo Fisher Scientific (Waltham, MA) Stealth RNAi siRNA, negative control 10620312-367123 D11/E02 Thermo Fisher Scientific (Waltham, MA) Stealth RNAi siRNAs, TRPV4 Sequence 1 RSS330372 RSS330373 RSS330374 Thermo Fisher Scientific (Waltham, MA) Sequence 2 Sequence 3 Product Catalog number Manufacturer .. DMEM D5796 Sigma-Aldrich (St. Louis, MO) FBS F2442 Sigma-Aldrich (St. Louis, MO) Penicillin/streptomycin 26253-84 Nacalai Tesque (Kyoto, Japan) Collagenase type 2 LS004176 Worthington Biochemical (Lakewood, NJ) Rat recombinant IL-1β 092-04263 Wako (Osaka, Japan) CCK-8 CK04 Dojindo Laboratories (Kumamoto, Japan) Model 680 microplate reader Bio-Rad (Hercules, CA) Staining Hematoxylin 30002 Muto (Tokyo, Japan) Eosin 058-00062 Wako (Osaka, Japan) Safranin-O S-0145 Tokyo Chemical Industry (Tokyo, Japan) Fast green 10720 Chroma Gesellschaft Schmidt (Munster, Germany) In situ cell death detection kit, fluorescein 11684795910 Roche (Basel, Switzerland) SA-β-gal staining kit 9860 Cell Signaling Technology (Danvers, MA) 4% Paraformaldehyde phosphate buffer solution 163-20145 Wako (Osaka, Japan) Alexa Fluor 488 secondary antibody A-21467 Thermo Fisher Scientific (Waltham, MA) Alexa Fluor 568 secondary antibody A-10042 Thermo Fisher Scientific (Waltham, MA) Alexa Fluor 647 secondary antibody A-31571 Thermo Fisher Scientific (Waltham, MA) Rat IgG Isotype Control 31933 Thermo Fisher Scientific (Waltham, MA) DAPI D1306 Thermo Fisher Scientific (Waltham, MA) BZ-X700 microscope Keyence (Osaka, Japan) Western blotting 3-(N-Morpholino)propanesulfonic acid 23438-64 Nacalai Tesque (Kyoto, Japan) Protease inhibitor cocktail 25955-11 Nacalai Tesque (Kyoto, Japan) Tris(hydroxymethyl)aminomethane 35434-21 Nacalai Tesque (Kyoto, Japan) Chemi-Lumi One Super 02230-30 Nacalai Tesque (Kyoto, Japan) Phosphatase inhibitor cocktails 2 P5726 Sigma-Aldrich (St. Louis, MO) Phosphatase inhibitor cocktails 3 P0044 Sigma-Aldrich (St. Louis, MO) Glycine 12-1210 Sigma-Aldrich (St. Louis, MO) T-PER tissue protein extraction reagent 78510 Thermo Fisher Scientific (Waltham, MA) Pierce BCA protein assay kit 23227 Thermo Fisher Scientific (Waltham, MA) 7.5%–15.0% Polyacrylamide gel SDG-581 Bio Craft (Tokyo, Japan) 4× Laemmli sample buffer 1610747 Bio-Rad (Hercules, CA) 2-Mercaptoethanol 21438-82 Nacalai Tesque (Kyoto, Japan) SDS 191-07145 Wako (Osaka, Japan) 0.2 PVDF Western blotting membrane 10600021 GE Healthcare (Chicago, IL) Anti-rabbit secondary antibody NA934 GE Healthcare (Chicago, IL) Product Catalog number Manufacturer

    Transfection:

    Article Title: Transient Receptor Potential Vanilloid 4 Knockdown Decreases Extracellular Matrix Synthesis via Autophagy Suppression in the Rat Intervertebral Disc
    Article Snippet: .. TRPV1 (rabbit, polyclonal) ACC-030 Almone labs (Jerusalem, Israel) TRPV4 (rabbit, polyclonal) ACC-124 Almone labs (Jerusalem, Israel) TRPV5 (rabbit, polyclonal) ACC-035 Almone labs (Jerusalem, Israel) AMPK (rabbit, polyclonal) 2532S Cell Signaling Technology (Danvers, MA) P-AMPK (rabbit, monoclonal) 2535S Cell Signaling Technology (Danvers, MA) mTOR (rabbit, monoclonal) 2983 Cell Signaling Technology (Danvers, MA) RAPTOR (rabbit, monoclonal) 2280 Cell Signaling Technology (Danvers, MA) RICTOR (rabbit, monoclonal) 2114 Cell Signaling Technology (Danvers, MA) p70/S6K (rabbit, monoclonal) 2708 Cell Signaling Technology (Danvers, MA) LC3 (rabbit, monoclonal) 3868 Cell Signaling Technology (Danvers, MA) p62/SQSTM1 (mouse, monoclonal) ab56416 Abcam (Cambridge, UK) Cleaved caspase-9 (rabbit, polyclonal) 9507 Cell Signaling Technology (Danvers, MA) PARP (rabbit, monoclonal) 9532 Cell Signaling Technology (Danvers, MA) Cleaved PARP (rabbit, monoclonal) 5625 Cell Signaling Technology (Danvers, MA) p53 (mouse, monoclonal) 2524 Cell Signaling Technology (Danvers, MA) p21/CIP1 (mouse, monoclonal) sc-6246 Santa Cruz Biotechnology (Santa Cruz, CA) Brachyury (mouse, monoclonal) sc-166962 Santa Cruz Biotechnology (Santa Cruz, CA) CD24 (mouse, monoclonal) sc-19585 Santa Cruz Biotechnology (Santa Cruz, CA) p16/INK4a (mouse, monoclonal) sc-1661 Santa Cruz Biotechnology (Santa Cruz, CA) COL2A1 (mouse, monoclonal) sc-52658 Santa Cruz Biotechnology (Santa Cruz, CA) Aggrecan (mouse, monoclonal) ab3778 Abcam (Cambridge, UK) MMP3 (rabbit, monoclonal) ab52915 Abcam (Cambridge, UK) MMP13 (rabbit, polyclonal) ab39012 Abcam (Cambridge, UK) TIMP1 (mouse, monoclonal) sc-21734 Santa Cruz Biotechnology (Santa Cruz, CA) TIMP2 (rabbit, monoclonal) 5738 Cell Signaling Technology (Danvers, MA) Tubulin (mouse, monoclonal) T9026 Sigma-Aldrich (St. Louis, MO) RNAi Lipofectamine RNAiMAX transfection reagent 13778150 Thermo Fisher Scientific (Waltham, MA) Invivofectamine 3.0 Reagent IVF3001 Thermo Fisher Scientific (Waltham, MA) Opti-minimal essential medium I 3198070 Thermo Fisher Scientific (Waltham, MA) Stealth RNAi siRNA, negative control 10620312-367123 D11/E02 Thermo Fisher Scientific (Waltham, MA) Stealth RNAi siRNAs, TRPV4 Sequence 1 RSS330372 RSS330373 RSS330374 Thermo Fisher Scientific (Waltham, MA) Sequence 2 Sequence 3 Product Catalog number Manufacturer .. DMEM D5796 Sigma-Aldrich (St. Louis, MO) FBS F2442 Sigma-Aldrich (St. Louis, MO) Penicillin/streptomycin 26253-84 Nacalai Tesque (Kyoto, Japan) Collagenase type 2 LS004176 Worthington Biochemical (Lakewood, NJ) Rat recombinant IL-1β 092-04263 Wako (Osaka, Japan) CCK-8 CK04 Dojindo Laboratories (Kumamoto, Japan) Model 680 microplate reader Bio-Rad (Hercules, CA) Staining Hematoxylin 30002 Muto (Tokyo, Japan) Eosin 058-00062 Wako (Osaka, Japan) Safranin-O S-0145 Tokyo Chemical Industry (Tokyo, Japan) Fast green 10720 Chroma Gesellschaft Schmidt (Munster, Germany) In situ cell death detection kit, fluorescein 11684795910 Roche (Basel, Switzerland) SA-β-gal staining kit 9860 Cell Signaling Technology (Danvers, MA) 4% Paraformaldehyde phosphate buffer solution 163-20145 Wako (Osaka, Japan) Alexa Fluor 488 secondary antibody A-21467 Thermo Fisher Scientific (Waltham, MA) Alexa Fluor 568 secondary antibody A-10042 Thermo Fisher Scientific (Waltham, MA) Alexa Fluor 647 secondary antibody A-31571 Thermo Fisher Scientific (Waltham, MA) Rat IgG Isotype Control 31933 Thermo Fisher Scientific (Waltham, MA) DAPI D1306 Thermo Fisher Scientific (Waltham, MA) BZ-X700 microscope Keyence (Osaka, Japan) Western blotting 3-(N-Morpholino)propanesulfonic acid 23438-64 Nacalai Tesque (Kyoto, Japan) Protease inhibitor cocktail 25955-11 Nacalai Tesque (Kyoto, Japan) Tris(hydroxymethyl)aminomethane 35434-21 Nacalai Tesque (Kyoto, Japan) Chemi-Lumi One Super 02230-30 Nacalai Tesque (Kyoto, Japan) Phosphatase inhibitor cocktails 2 P5726 Sigma-Aldrich (St. Louis, MO) Phosphatase inhibitor cocktails 3 P0044 Sigma-Aldrich (St. Louis, MO) Glycine 12-1210 Sigma-Aldrich (St. Louis, MO) T-PER tissue protein extraction reagent 78510 Thermo Fisher Scientific (Waltham, MA) Pierce BCA protein assay kit 23227 Thermo Fisher Scientific (Waltham, MA) 7.5%–15.0% Polyacrylamide gel SDG-581 Bio Craft (Tokyo, Japan) 4× Laemmli sample buffer 1610747 Bio-Rad (Hercules, CA) 2-Mercaptoethanol 21438-82 Nacalai Tesque (Kyoto, Japan) SDS 191-07145 Wako (Osaka, Japan) 0.2 PVDF Western blotting membrane 10600021 GE Healthcare (Chicago, IL) Anti-rabbit secondary antibody NA934 GE Healthcare (Chicago, IL) Product Catalog number Manufacturer

    Negative Control:

    Article Title: Transient Receptor Potential Vanilloid 4 Knockdown Decreases Extracellular Matrix Synthesis via Autophagy Suppression in the Rat Intervertebral Disc
    Article Snippet: .. TRPV1 (rabbit, polyclonal) ACC-030 Almone labs (Jerusalem, Israel) TRPV4 (rabbit, polyclonal) ACC-124 Almone labs (Jerusalem, Israel) TRPV5 (rabbit, polyclonal) ACC-035 Almone labs (Jerusalem, Israel) AMPK (rabbit, polyclonal) 2532S Cell Signaling Technology (Danvers, MA) P-AMPK (rabbit, monoclonal) 2535S Cell Signaling Technology (Danvers, MA) mTOR (rabbit, monoclonal) 2983 Cell Signaling Technology (Danvers, MA) RAPTOR (rabbit, monoclonal) 2280 Cell Signaling Technology (Danvers, MA) RICTOR (rabbit, monoclonal) 2114 Cell Signaling Technology (Danvers, MA) p70/S6K (rabbit, monoclonal) 2708 Cell Signaling Technology (Danvers, MA) LC3 (rabbit, monoclonal) 3868 Cell Signaling Technology (Danvers, MA) p62/SQSTM1 (mouse, monoclonal) ab56416 Abcam (Cambridge, UK) Cleaved caspase-9 (rabbit, polyclonal) 9507 Cell Signaling Technology (Danvers, MA) PARP (rabbit, monoclonal) 9532 Cell Signaling Technology (Danvers, MA) Cleaved PARP (rabbit, monoclonal) 5625 Cell Signaling Technology (Danvers, MA) p53 (mouse, monoclonal) 2524 Cell Signaling Technology (Danvers, MA) p21/CIP1 (mouse, monoclonal) sc-6246 Santa Cruz Biotechnology (Santa Cruz, CA) Brachyury (mouse, monoclonal) sc-166962 Santa Cruz Biotechnology (Santa Cruz, CA) CD24 (mouse, monoclonal) sc-19585 Santa Cruz Biotechnology (Santa Cruz, CA) p16/INK4a (mouse, monoclonal) sc-1661 Santa Cruz Biotechnology (Santa Cruz, CA) COL2A1 (mouse, monoclonal) sc-52658 Santa Cruz Biotechnology (Santa Cruz, CA) Aggrecan (mouse, monoclonal) ab3778 Abcam (Cambridge, UK) MMP3 (rabbit, monoclonal) ab52915 Abcam (Cambridge, UK) MMP13 (rabbit, polyclonal) ab39012 Abcam (Cambridge, UK) TIMP1 (mouse, monoclonal) sc-21734 Santa Cruz Biotechnology (Santa Cruz, CA) TIMP2 (rabbit, monoclonal) 5738 Cell Signaling Technology (Danvers, MA) Tubulin (mouse, monoclonal) T9026 Sigma-Aldrich (St. Louis, MO) RNAi Lipofectamine RNAiMAX transfection reagent 13778150 Thermo Fisher Scientific (Waltham, MA) Invivofectamine 3.0 Reagent IVF3001 Thermo Fisher Scientific (Waltham, MA) Opti-minimal essential medium I 3198070 Thermo Fisher Scientific (Waltham, MA) Stealth RNAi siRNA, negative control 10620312-367123 D11/E02 Thermo Fisher Scientific (Waltham, MA) Stealth RNAi siRNAs, TRPV4 Sequence 1 RSS330372 RSS330373 RSS330374 Thermo Fisher Scientific (Waltham, MA) Sequence 2 Sequence 3 Product Catalog number Manufacturer .. DMEM D5796 Sigma-Aldrich (St. Louis, MO) FBS F2442 Sigma-Aldrich (St. Louis, MO) Penicillin/streptomycin 26253-84 Nacalai Tesque (Kyoto, Japan) Collagenase type 2 LS004176 Worthington Biochemical (Lakewood, NJ) Rat recombinant IL-1β 092-04263 Wako (Osaka, Japan) CCK-8 CK04 Dojindo Laboratories (Kumamoto, Japan) Model 680 microplate reader Bio-Rad (Hercules, CA) Staining Hematoxylin 30002 Muto (Tokyo, Japan) Eosin 058-00062 Wako (Osaka, Japan) Safranin-O S-0145 Tokyo Chemical Industry (Tokyo, Japan) Fast green 10720 Chroma Gesellschaft Schmidt (Munster, Germany) In situ cell death detection kit, fluorescein 11684795910 Roche (Basel, Switzerland) SA-β-gal staining kit 9860 Cell Signaling Technology (Danvers, MA) 4% Paraformaldehyde phosphate buffer solution 163-20145 Wako (Osaka, Japan) Alexa Fluor 488 secondary antibody A-21467 Thermo Fisher Scientific (Waltham, MA) Alexa Fluor 568 secondary antibody A-10042 Thermo Fisher Scientific (Waltham, MA) Alexa Fluor 647 secondary antibody A-31571 Thermo Fisher Scientific (Waltham, MA) Rat IgG Isotype Control 31933 Thermo Fisher Scientific (Waltham, MA) DAPI D1306 Thermo Fisher Scientific (Waltham, MA) BZ-X700 microscope Keyence (Osaka, Japan) Western blotting 3-(N-Morpholino)propanesulfonic acid 23438-64 Nacalai Tesque (Kyoto, Japan) Protease inhibitor cocktail 25955-11 Nacalai Tesque (Kyoto, Japan) Tris(hydroxymethyl)aminomethane 35434-21 Nacalai Tesque (Kyoto, Japan) Chemi-Lumi One Super 02230-30 Nacalai Tesque (Kyoto, Japan) Phosphatase inhibitor cocktails 2 P5726 Sigma-Aldrich (St. Louis, MO) Phosphatase inhibitor cocktails 3 P0044 Sigma-Aldrich (St. Louis, MO) Glycine 12-1210 Sigma-Aldrich (St. Louis, MO) T-PER tissue protein extraction reagent 78510 Thermo Fisher Scientific (Waltham, MA) Pierce BCA protein assay kit 23227 Thermo Fisher Scientific (Waltham, MA) 7.5%–15.0% Polyacrylamide gel SDG-581 Bio Craft (Tokyo, Japan) 4× Laemmli sample buffer 1610747 Bio-Rad (Hercules, CA) 2-Mercaptoethanol 21438-82 Nacalai Tesque (Kyoto, Japan) SDS 191-07145 Wako (Osaka, Japan) 0.2 PVDF Western blotting membrane 10600021 GE Healthcare (Chicago, IL) Anti-rabbit secondary antibody NA934 GE Healthcare (Chicago, IL) Product Catalog number Manufacturer

    Sequencing:

    Article Title: Transient Receptor Potential Vanilloid 4 Knockdown Decreases Extracellular Matrix Synthesis via Autophagy Suppression in the Rat Intervertebral Disc
    Article Snippet: .. TRPV1 (rabbit, polyclonal) ACC-030 Almone labs (Jerusalem, Israel) TRPV4 (rabbit, polyclonal) ACC-124 Almone labs (Jerusalem, Israel) TRPV5 (rabbit, polyclonal) ACC-035 Almone labs (Jerusalem, Israel) AMPK (rabbit, polyclonal) 2532S Cell Signaling Technology (Danvers, MA) P-AMPK (rabbit, monoclonal) 2535S Cell Signaling Technology (Danvers, MA) mTOR (rabbit, monoclonal) 2983 Cell Signaling Technology (Danvers, MA) RAPTOR (rabbit, monoclonal) 2280 Cell Signaling Technology (Danvers, MA) RICTOR (rabbit, monoclonal) 2114 Cell Signaling Technology (Danvers, MA) p70/S6K (rabbit, monoclonal) 2708 Cell Signaling Technology (Danvers, MA) LC3 (rabbit, monoclonal) 3868 Cell Signaling Technology (Danvers, MA) p62/SQSTM1 (mouse, monoclonal) ab56416 Abcam (Cambridge, UK) Cleaved caspase-9 (rabbit, polyclonal) 9507 Cell Signaling Technology (Danvers, MA) PARP (rabbit, monoclonal) 9532 Cell Signaling Technology (Danvers, MA) Cleaved PARP (rabbit, monoclonal) 5625 Cell Signaling Technology (Danvers, MA) p53 (mouse, monoclonal) 2524 Cell Signaling Technology (Danvers, MA) p21/CIP1 (mouse, monoclonal) sc-6246 Santa Cruz Biotechnology (Santa Cruz, CA) Brachyury (mouse, monoclonal) sc-166962 Santa Cruz Biotechnology (Santa Cruz, CA) CD24 (mouse, monoclonal) sc-19585 Santa Cruz Biotechnology (Santa Cruz, CA) p16/INK4a (mouse, monoclonal) sc-1661 Santa Cruz Biotechnology (Santa Cruz, CA) COL2A1 (mouse, monoclonal) sc-52658 Santa Cruz Biotechnology (Santa Cruz, CA) Aggrecan (mouse, monoclonal) ab3778 Abcam (Cambridge, UK) MMP3 (rabbit, monoclonal) ab52915 Abcam (Cambridge, UK) MMP13 (rabbit, polyclonal) ab39012 Abcam (Cambridge, UK) TIMP1 (mouse, monoclonal) sc-21734 Santa Cruz Biotechnology (Santa Cruz, CA) TIMP2 (rabbit, monoclonal) 5738 Cell Signaling Technology (Danvers, MA) Tubulin (mouse, monoclonal) T9026 Sigma-Aldrich (St. Louis, MO) RNAi Lipofectamine RNAiMAX transfection reagent 13778150 Thermo Fisher Scientific (Waltham, MA) Invivofectamine 3.0 Reagent IVF3001 Thermo Fisher Scientific (Waltham, MA) Opti-minimal essential medium I 3198070 Thermo Fisher Scientific (Waltham, MA) Stealth RNAi siRNA, negative control 10620312-367123 D11/E02 Thermo Fisher Scientific (Waltham, MA) Stealth RNAi siRNAs, TRPV4 Sequence 1 RSS330372 RSS330373 RSS330374 Thermo Fisher Scientific (Waltham, MA) Sequence 2 Sequence 3 Product Catalog number Manufacturer .. DMEM D5796 Sigma-Aldrich (St. Louis, MO) FBS F2442 Sigma-Aldrich (St. Louis, MO) Penicillin/streptomycin 26253-84 Nacalai Tesque (Kyoto, Japan) Collagenase type 2 LS004176 Worthington Biochemical (Lakewood, NJ) Rat recombinant IL-1β 092-04263 Wako (Osaka, Japan) CCK-8 CK04 Dojindo Laboratories (Kumamoto, Japan) Model 680 microplate reader Bio-Rad (Hercules, CA) Staining Hematoxylin 30002 Muto (Tokyo, Japan) Eosin 058-00062 Wako (Osaka, Japan) Safranin-O S-0145 Tokyo Chemical Industry (Tokyo, Japan) Fast green 10720 Chroma Gesellschaft Schmidt (Munster, Germany) In situ cell death detection kit, fluorescein 11684795910 Roche (Basel, Switzerland) SA-β-gal staining kit 9860 Cell Signaling Technology (Danvers, MA) 4% Paraformaldehyde phosphate buffer solution 163-20145 Wako (Osaka, Japan) Alexa Fluor 488 secondary antibody A-21467 Thermo Fisher Scientific (Waltham, MA) Alexa Fluor 568 secondary antibody A-10042 Thermo Fisher Scientific (Waltham, MA) Alexa Fluor 647 secondary antibody A-31571 Thermo Fisher Scientific (Waltham, MA) Rat IgG Isotype Control 31933 Thermo Fisher Scientific (Waltham, MA) DAPI D1306 Thermo Fisher Scientific (Waltham, MA) BZ-X700 microscope Keyence (Osaka, Japan) Western blotting 3-(N-Morpholino)propanesulfonic acid 23438-64 Nacalai Tesque (Kyoto, Japan) Protease inhibitor cocktail 25955-11 Nacalai Tesque (Kyoto, Japan) Tris(hydroxymethyl)aminomethane 35434-21 Nacalai Tesque (Kyoto, Japan) Chemi-Lumi One Super 02230-30 Nacalai Tesque (Kyoto, Japan) Phosphatase inhibitor cocktails 2 P5726 Sigma-Aldrich (St. Louis, MO) Phosphatase inhibitor cocktails 3 P0044 Sigma-Aldrich (St. Louis, MO) Glycine 12-1210 Sigma-Aldrich (St. Louis, MO) T-PER tissue protein extraction reagent 78510 Thermo Fisher Scientific (Waltham, MA) Pierce BCA protein assay kit 23227 Thermo Fisher Scientific (Waltham, MA) 7.5%–15.0% Polyacrylamide gel SDG-581 Bio Craft (Tokyo, Japan) 4× Laemmli sample buffer 1610747 Bio-Rad (Hercules, CA) 2-Mercaptoethanol 21438-82 Nacalai Tesque (Kyoto, Japan) SDS 191-07145 Wako (Osaka, Japan) 0.2 PVDF Western blotting membrane 10600021 GE Healthcare (Chicago, IL) Anti-rabbit secondary antibody NA934 GE Healthcare (Chicago, IL) Product Catalog number Manufacturer

    Retroviral:

    Article Title: Human Papillomavirus Infection Blunts STING Signaling and Alters Downstream Response
    Article Snippet: .. Retroviral expression was confirmed by western blotting using antibodies for p16 INK4A (Santa Cruz 56330 clone JC8) and PTPN14 (Cell Signaling 13808 clone D5T6Y), key markers of E6 and E7 expression. .. For western blot analysis, cells were lysed in 1x high SDS RIPA protein lysis buffer (50 mM Tris-HCl pH 8.0, 150 mM NaCl, 1% NP40, 0.5% sodium deoxycholate, 0.5% SDS), supplemented with 1x protease inhibitor cocktail (Sigma P1860), 1mM PMSF and 1x PhosSTOP phosphatase inhibitor cocktail (Roche 04906845001).

    Expressing:

    Article Title: Human Papillomavirus Infection Blunts STING Signaling and Alters Downstream Response
    Article Snippet: .. Retroviral expression was confirmed by western blotting using antibodies for p16 INK4A (Santa Cruz 56330 clone JC8) and PTPN14 (Cell Signaling 13808 clone D5T6Y), key markers of E6 and E7 expression. .. For western blot analysis, cells were lysed in 1x high SDS RIPA protein lysis buffer (50 mM Tris-HCl pH 8.0, 150 mM NaCl, 1% NP40, 0.5% sodium deoxycholate, 0.5% SDS), supplemented with 1x protease inhibitor cocktail (Sigma P1860), 1mM PMSF and 1x PhosSTOP phosphatase inhibitor cocktail (Roche 04906845001).

    Western Blot:

    Article Title: Human Papillomavirus Infection Blunts STING Signaling and Alters Downstream Response
    Article Snippet: .. Retroviral expression was confirmed by western blotting using antibodies for p16 INK4A (Santa Cruz 56330 clone JC8) and PTPN14 (Cell Signaling 13808 clone D5T6Y), key markers of E6 and E7 expression. .. For western blot analysis, cells were lysed in 1x high SDS RIPA protein lysis buffer (50 mM Tris-HCl pH 8.0, 150 mM NaCl, 1% NP40, 0.5% sodium deoxycholate, 0.5% SDS), supplemented with 1x protease inhibitor cocktail (Sigma P1860), 1mM PMSF and 1x PhosSTOP phosphatase inhibitor cocktail (Roche 04906845001).



    Similar Products

    p16 ink4a  (Bioss)
    94
    Bioss p16 ink4a
    CI in older and D‐gal‐induced mice is accompanied by microglial senescence and elevated ghrelin levels. (A) Experimental design of the D‐gal‐induced aging mouse model. (B) Percentage change in body weight of mice in the Young, Old, Control, and D‐gal groups, normalized to baseline body weight at the start of the experiment. (C) The NOL test comparing the mice from the Young and Old groups and from the Control and D‐gal groups. The recognition index was calculated as the ratio of time spent exploring in the novel location to the total time spent exploring both locations. (D) NOR test comparing the mice from the Young and Old groups and from the Control and D‐gal groups. The recognition index was calculated as the ratio of time spent exploring the novel object relative to the total exploration time for both objects. (E) MWM test comparing the mice from the Young and Old groups and from the Control and D‐gal groups, including swimming trajectories and escape latency. (F) Western blot analysis of senescence markers <t>p16</t> <t>Ink4a</t> , p21 Cip1 , LaminB1, γH2AX, and HMGB1 in the mice brain tissue from the Young, Old, Control, and D‐gal groups. (G) Immunofluorescence co‐localization of Iba‐1 + microglia with LaminB1, γH2AX, and HMGB1 in the hippocampal CA1 and CA3 regions of the mice from Young, Old, Control, and D‐gal groups (magnification: ×400, scale bar = 25 μm). (H) ELISA‐based quantification of hippocampal cytokines (IL‐1α, IL‐1β, G‐CSF, IL‐4, IL‐6, IL‐8, CXCL10, LIF, MCP1, and M‐CSF) in the Young, Old, Control, and D‐gal groups. (I) ELISA measurement of plasma total ghrelin and acylated ghrelin in the Young, Old, Control, and D‐gal groups. ** p < 0.01, *** p < 0.001.
    P16 Ink4a, supplied by Bioss, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/p16+ink4a/CDKN2A+p16-INK4a+Polyclonal+Antibody/pmc13621234-152-34-38
    Average 94 stars, based on 1 article reviews
    p16 ink4a - by Bioz Stars, 2026-10
    94/100 stars
    		  Buy from Supplier
    p16  (Proteintech)
    96
    Proteintech p16
    CI in older and D‐gal‐induced mice is accompanied by microglial senescence and elevated ghrelin levels. (A) Experimental design of the D‐gal‐induced aging mouse model. (B) Percentage change in body weight of mice in the Young, Old, Control, and D‐gal groups, normalized to baseline body weight at the start of the experiment. (C) The NOL test comparing the mice from the Young and Old groups and from the Control and D‐gal groups. The recognition index was calculated as the ratio of time spent exploring in the novel location to the total time spent exploring both locations. (D) NOR test comparing the mice from the Young and Old groups and from the Control and D‐gal groups. The recognition index was calculated as the ratio of time spent exploring the novel object relative to the total exploration time for both objects. (E) MWM test comparing the mice from the Young and Old groups and from the Control and D‐gal groups, including swimming trajectories and escape latency. (F) Western blot analysis of senescence markers <t>p16</t> <t>Ink4a</t> , p21 Cip1 , LaminB1, γH2AX, and HMGB1 in the mice brain tissue from the Young, Old, Control, and D‐gal groups. (G) Immunofluorescence co‐localization of Iba‐1 + microglia with LaminB1, γH2AX, and HMGB1 in the hippocampal CA1 and CA3 regions of the mice from Young, Old, Control, and D‐gal groups (magnification: ×400, scale bar = 25 μm). (H) ELISA‐based quantification of hippocampal cytokines (IL‐1α, IL‐1β, G‐CSF, IL‐4, IL‐6, IL‐8, CXCL10, LIF, MCP1, and M‐CSF) in the Young, Old, Control, and D‐gal groups. (I) ELISA measurement of plasma total ghrelin and acylated ghrelin in the Young, Old, Control, and D‐gal groups. ** p < 0.01, *** p < 0.001.
    P16, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/p16+ink4a/P16-INK4A+Antibody/pm42789189-98-75-78
    Average 96 stars, based on 1 article reviews
    p16 - by Bioz Stars, 2026-10
    96/100 stars
    		  Buy from Supplier
    rabbit anti p16  (Proteintech)
    96
    Proteintech rabbit anti p16
    CI in older and D‐gal‐induced mice is accompanied by microglial senescence and elevated ghrelin levels. (A) Experimental design of the D‐gal‐induced aging mouse model. (B) Percentage change in body weight of mice in the Young, Old, Control, and D‐gal groups, normalized to baseline body weight at the start of the experiment. (C) The NOL test comparing the mice from the Young and Old groups and from the Control and D‐gal groups. The recognition index was calculated as the ratio of time spent exploring in the novel location to the total time spent exploring both locations. (D) NOR test comparing the mice from the Young and Old groups and from the Control and D‐gal groups. The recognition index was calculated as the ratio of time spent exploring the novel object relative to the total exploration time for both objects. (E) MWM test comparing the mice from the Young and Old groups and from the Control and D‐gal groups, including swimming trajectories and escape latency. (F) Western blot analysis of senescence markers <t>p16</t> <t>Ink4a</t> , p21 Cip1 , LaminB1, γH2AX, and HMGB1 in the mice brain tissue from the Young, Old, Control, and D‐gal groups. (G) Immunofluorescence co‐localization of Iba‐1 + microglia with LaminB1, γH2AX, and HMGB1 in the hippocampal CA1 and CA3 regions of the mice from Young, Old, Control, and D‐gal groups (magnification: ×400, scale bar = 25 μm). (H) ELISA‐based quantification of hippocampal cytokines (IL‐1α, IL‐1β, G‐CSF, IL‐4, IL‐6, IL‐8, CXCL10, LIF, MCP1, and M‐CSF) in the Young, Old, Control, and D‐gal groups. (I) ELISA measurement of plasma total ghrelin and acylated ghrelin in the Young, Old, Control, and D‐gal groups. ** p < 0.01, *** p < 0.001.
    Rabbit Anti P16, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/p16+ink4a/P16-INK4A+Fusion+Protein/pmc13628733-104-18-20
    Average 96 stars, based on 1 article reviews
    rabbit anti p16 - by Bioz Stars, 2026-10
    96/100 stars
    		  Buy from Supplier
    p16 ink4a mrna  (Miltenyi Biotec)
    90
    Miltenyi Biotec p16 ink4a mrna
    CI in older and D‐gal‐induced mice is accompanied by microglial senescence and elevated ghrelin levels. (A) Experimental design of the D‐gal‐induced aging mouse model. (B) Percentage change in body weight of mice in the Young, Old, Control, and D‐gal groups, normalized to baseline body weight at the start of the experiment. (C) The NOL test comparing the mice from the Young and Old groups and from the Control and D‐gal groups. The recognition index was calculated as the ratio of time spent exploring in the novel location to the total time spent exploring both locations. (D) NOR test comparing the mice from the Young and Old groups and from the Control and D‐gal groups. The recognition index was calculated as the ratio of time spent exploring the novel object relative to the total exploration time for both objects. (E) MWM test comparing the mice from the Young and Old groups and from the Control and D‐gal groups, including swimming trajectories and escape latency. (F) Western blot analysis of senescence markers <t>p16</t> <t>Ink4a</t> , p21 Cip1 , LaminB1, γH2AX, and HMGB1 in the mice brain tissue from the Young, Old, Control, and D‐gal groups. (G) Immunofluorescence co‐localization of Iba‐1 + microglia with LaminB1, γH2AX, and HMGB1 in the hippocampal CA1 and CA3 regions of the mice from Young, Old, Control, and D‐gal groups (magnification: ×400, scale bar = 25 μm). (H) ELISA‐based quantification of hippocampal cytokines (IL‐1α, IL‐1β, G‐CSF, IL‐4, IL‐6, IL‐8, CXCL10, LIF, MCP1, and M‐CSF) in the Young, Old, Control, and D‐gal groups. (I) ELISA measurement of plasma total ghrelin and acylated ghrelin in the Young, Old, Control, and D‐gal groups. ** p < 0.01, *** p < 0.001.
    P16 Ink4a Mrna, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/p16+ink4a/p16+Antibody%2C+anti-human%2C+REAfinity/pmc13123506-163-31-28
    Average 90 stars, based on 1 article reviews
    p16 ink4a mrna - by Bioz Stars, 2026-10
    90/100 stars
    		  Buy from Supplier
    primary antibodies against p16  (Bioss)
    94
    Bioss primary antibodies against p16
    CI in older and D‐gal‐induced mice is accompanied by microglial senescence and elevated ghrelin levels. (A) Experimental design of the D‐gal‐induced aging mouse model. (B) Percentage change in body weight of mice in the Young, Old, Control, and D‐gal groups, normalized to baseline body weight at the start of the experiment. (C) The NOL test comparing the mice from the Young and Old groups and from the Control and D‐gal groups. The recognition index was calculated as the ratio of time spent exploring in the novel location to the total time spent exploring both locations. (D) NOR test comparing the mice from the Young and Old groups and from the Control and D‐gal groups. The recognition index was calculated as the ratio of time spent exploring the novel object relative to the total exploration time for both objects. (E) MWM test comparing the mice from the Young and Old groups and from the Control and D‐gal groups, including swimming trajectories and escape latency. (F) Western blot analysis of senescence markers <t>p16</t> <t>Ink4a</t> , p21 Cip1 , LaminB1, γH2AX, and HMGB1 in the mice brain tissue from the Young, Old, Control, and D‐gal groups. (G) Immunofluorescence co‐localization of Iba‐1 + microglia with LaminB1, γH2AX, and HMGB1 in the hippocampal CA1 and CA3 regions of the mice from Young, Old, Control, and D‐gal groups (magnification: ×400, scale bar = 25 μm). (H) ELISA‐based quantification of hippocampal cytokines (IL‐1α, IL‐1β, G‐CSF, IL‐4, IL‐6, IL‐8, CXCL10, LIF, MCP1, and M‐CSF) in the Young, Old, Control, and D‐gal groups. (I) ELISA measurement of plasma total ghrelin and acylated ghrelin in the Young, Old, Control, and D‐gal groups. ** p < 0.01, *** p < 0.001.
    Primary Antibodies Against P16, supplied by Bioss, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/p16+ink4a/CDKN2A+p16-INK4a+Polyclonal+Antibody%2C+ALEXA+FLUOR+488+Conjugated/10__3390_slash_ijms27083618-185-0-6
    Average 94 stars, based on 1 article reviews
    primary antibodies against p16 - by Bioz Stars, 2026-10
    94/100 stars
    		  Buy from Supplier
    rabbit anti p16  (Cell Signaling Technology Inc)
    96
    Cell Signaling Technology Inc rabbit anti p16
    Senescence was detected more frequently in MTJ-specific and vascular endothelial cells compared to the overall cell population. Representative in situ hybridisation images of young and old mouse MTJ (n = 4 per age group) showing senescence markers ( a ) <t>p16</t> and ( b ) p21.Red: Col22 (MTJ marker); yellow: ( a ) p16 and ( b ) p21; cyan: VWF (endothelial cell marker); grey: DAPI. Blue DAPI image on the right shows the interface between the muscle and tendon indicated by orange dashed lines. Scale bar is 50 µm. Percentage of ( c ) p16 and ( d ) p21 positive cells were measured in Col22-positive MTJ-specific cells, VWF-positive endothelial cells, and all cells in the field of view. Y: young; O: old. Two-way ANOVA followed by Tukey’s multiple comparisons test was used to calculate the significance between the age groups and cell populations. Data are presented as mean ± SD: *p < 0.05 **p < 0.01 ****p < 0.0001
    Rabbit Anti P16, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/p16+ink4a/p16+INK4A+Rabbit+mAb/pmc13050345-89-24-28
    Average 96 stars, based on 1 article reviews
    rabbit anti p16 - by Bioz Stars, 2026-10
    96/100 stars
    		  Buy from Supplier
    p16 ink4a antibody f 12  (Santa Cruz Biotechnology)
    96
    Santa Cruz Biotechnology p16 ink4a antibody f 12
    M6A modifications within total mRNA were significantly enhanced during the aging process . ( A ) Left: representative photographs of 2-month and 20-month-old male C57BL/6 mouse; Right: representative photographs of SA-β-gal staining in kidney tissues from young and old C57-BL/6 mice. ( B ) Left: representative photographs of SA-β-gal staining of P3 and P9 MEF cells; Right: the <t>p16</t> expression levels in P3 and P9 MEF cells. ( C ) Left: representative photographs of SA-β-gal staining of H 2 O 2 -induced premature senescence of NIH/3T3 cells; Right: the p16 expression levels in H 2 O 2 treated NIH/3T3 cells. ( D ) Poly(A)+ RNA was extracted from the brain and liver tissue of young and old C57-BL/6 mice and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( E ) Poly(A)+ RNA was extracted from the P3 and P9 MEF cells and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( F ) Poly(A)+ RNA was extracted from the P3 and P9 MEF cells and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( G ) Expression levels of m6A modification associated proteins were measured in P3 and P9 MEF cells, and the quantitative results were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; One sample t test (MEF P9 vs 1, METTL3: p =0.0027, t = 9.176).
    P16 Ink4a Antibody F 12, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/p16+ink4a/p16+Antibody/pmc12834426-37-42-47
    Average 96 stars, based on 1 article reviews
    p16 ink4a antibody f 12 - by Bioz Stars, 2026-10
    96/100 stars
    		  Buy from Supplier

    Image Search Results


    CI in older and D‐gal‐induced mice is accompanied by microglial senescence and elevated ghrelin levels. (A) Experimental design of the D‐gal‐induced aging mouse model. (B) Percentage change in body weight of mice in the Young, Old, Control, and D‐gal groups, normalized to baseline body weight at the start of the experiment. (C) The NOL test comparing the mice from the Young and Old groups and from the Control and D‐gal groups. The recognition index was calculated as the ratio of time spent exploring in the novel location to the total time spent exploring both locations. (D) NOR test comparing the mice from the Young and Old groups and from the Control and D‐gal groups. The recognition index was calculated as the ratio of time spent exploring the novel object relative to the total exploration time for both objects. (E) MWM test comparing the mice from the Young and Old groups and from the Control and D‐gal groups, including swimming trajectories and escape latency. (F) Western blot analysis of senescence markers p16 Ink4a , p21 Cip1 , LaminB1, γH2AX, and HMGB1 in the mice brain tissue from the Young, Old, Control, and D‐gal groups. (G) Immunofluorescence co‐localization of Iba‐1 + microglia with LaminB1, γH2AX, and HMGB1 in the hippocampal CA1 and CA3 regions of the mice from Young, Old, Control, and D‐gal groups (magnification: ×400, scale bar = 25 μm). (H) ELISA‐based quantification of hippocampal cytokines (IL‐1α, IL‐1β, G‐CSF, IL‐4, IL‐6, IL‐8, CXCL10, LIF, MCP1, and M‐CSF) in the Young, Old, Control, and D‐gal groups. (I) ELISA measurement of plasma total ghrelin and acylated ghrelin in the Young, Old, Control, and D‐gal groups. ** p < 0.01, *** p < 0.001.

    Journal: Aging Cell

    Article Title: Ghrelin Alleviates Aging‐Related Cognitive Impairment by Regulating Autophagy‐Related Signaling via the SHBG / JNK1 /Beclin1 Axis in Microglia

    doi: 10.1111/acel.70732

    Figure Lengend Snippet: CI in older and D‐gal‐induced mice is accompanied by microglial senescence and elevated ghrelin levels. (A) Experimental design of the D‐gal‐induced aging mouse model. (B) Percentage change in body weight of mice in the Young, Old, Control, and D‐gal groups, normalized to baseline body weight at the start of the experiment. (C) The NOL test comparing the mice from the Young and Old groups and from the Control and D‐gal groups. The recognition index was calculated as the ratio of time spent exploring in the novel location to the total time spent exploring both locations. (D) NOR test comparing the mice from the Young and Old groups and from the Control and D‐gal groups. The recognition index was calculated as the ratio of time spent exploring the novel object relative to the total exploration time for both objects. (E) MWM test comparing the mice from the Young and Old groups and from the Control and D‐gal groups, including swimming trajectories and escape latency. (F) Western blot analysis of senescence markers p16 Ink4a , p21 Cip1 , LaminB1, γH2AX, and HMGB1 in the mice brain tissue from the Young, Old, Control, and D‐gal groups. (G) Immunofluorescence co‐localization of Iba‐1 + microglia with LaminB1, γH2AX, and HMGB1 in the hippocampal CA1 and CA3 regions of the mice from Young, Old, Control, and D‐gal groups (magnification: ×400, scale bar = 25 μm). (H) ELISA‐based quantification of hippocampal cytokines (IL‐1α, IL‐1β, G‐CSF, IL‐4, IL‐6, IL‐8, CXCL10, LIF, MCP1, and M‐CSF) in the Young, Old, Control, and D‐gal groups. (I) ELISA measurement of plasma total ghrelin and acylated ghrelin in the Young, Old, Control, and D‐gal groups. ** p < 0.01, *** p < 0.001.

    Article Snippet: Following antibody stripping and re‐blocking, sections were incubated overnight at 4°C with a second primary antibody: γH2AX (1:400; ab81299, Abcam), high‐mobility group box 1 (HMGB1) (1:400; AWA10032 , Abiowell), p21 Cip1 (1:200; ab188224, Abcam), p16 Ink4a (1:200; bs‐23797R, Bioss, China), or Lamin B1 (1:400; ab8982, Abcam).

    Techniques: Control, Western Blot, Analysis, Tissue, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Clinical Proteomics

    Ghrelin administration alleviates D‐gal‐induced microglial senescence and CI. (A) Experimental design of ghrelin treatment in the D‐gal‐induced aging mouse model. (B) Percentage change in body weight of mice in the Control, Control + Ghrelin, D‐gal, and D‐gal + Ghrelin groups, normalized to baseline body weight at the start of the experiment. (C) The NOL test comparing the mice from the Control, Control + Ghrelin, D‐gal, and D‐gal + Ghrelin groups. The recognition index was calculated as the ratio of time spent exploring in the novel location to the total time spent exploring both locations. (D) NOR test comparing the mice from the Control, Control + Ghrelin, D‐gal, and D‐gal+Ghrelin groups. The recognition index was calculated as the ratio of time spent exploring the novel object relative to the total exploration time for both objects. (E) MWM test comparing the mice from the Control, Control + Ghrelin, D‐gal, and D‐gal + Ghrelin groups, including swimming trajectories and escape latency. (F) Western blot analysis of senescence markers p16 Ink4a , p21 Cip1 , LaminB1, γH2AX, and HMGB1 in the mice brain tissue from the Control, Control+Ghrelin, D‐gal, and D‐gal+Ghrelin groups. (G) Immunofluorescence co‐localization of Iba‐1 + microglia with LaminB1, γH2AX, and HMGB1 in the hippocampal CA1 region of the mice from Control, Control+Ghrelin, D‐gal, and D‐gal + Ghrelin groups (magnification: ×400, scale bar = 25 μm). (H) ELISA‐based quantification of hippocampal cytokines (IL‐1α, IL‐1β, G‐CSF, IL‐4, IL‐6, IL‐8, IP10/CXCL10, LIF, MCP1, and M‐CSF) in the Control, Control + Ghrelin, D‐gal, and D‐gal + Ghrelin groups. *** p < 0.001.

    Journal: Aging Cell

    Article Title: Ghrelin Alleviates Aging‐Related Cognitive Impairment by Regulating Autophagy‐Related Signaling via the SHBG / JNK1 /Beclin1 Axis in Microglia

    doi: 10.1111/acel.70732

    Figure Lengend Snippet: Ghrelin administration alleviates D‐gal‐induced microglial senescence and CI. (A) Experimental design of ghrelin treatment in the D‐gal‐induced aging mouse model. (B) Percentage change in body weight of mice in the Control, Control + Ghrelin, D‐gal, and D‐gal + Ghrelin groups, normalized to baseline body weight at the start of the experiment. (C) The NOL test comparing the mice from the Control, Control + Ghrelin, D‐gal, and D‐gal + Ghrelin groups. The recognition index was calculated as the ratio of time spent exploring in the novel location to the total time spent exploring both locations. (D) NOR test comparing the mice from the Control, Control + Ghrelin, D‐gal, and D‐gal+Ghrelin groups. The recognition index was calculated as the ratio of time spent exploring the novel object relative to the total exploration time for both objects. (E) MWM test comparing the mice from the Control, Control + Ghrelin, D‐gal, and D‐gal + Ghrelin groups, including swimming trajectories and escape latency. (F) Western blot analysis of senescence markers p16 Ink4a , p21 Cip1 , LaminB1, γH2AX, and HMGB1 in the mice brain tissue from the Control, Control+Ghrelin, D‐gal, and D‐gal+Ghrelin groups. (G) Immunofluorescence co‐localization of Iba‐1 + microglia with LaminB1, γH2AX, and HMGB1 in the hippocampal CA1 region of the mice from Control, Control+Ghrelin, D‐gal, and D‐gal + Ghrelin groups (magnification: ×400, scale bar = 25 μm). (H) ELISA‐based quantification of hippocampal cytokines (IL‐1α, IL‐1β, G‐CSF, IL‐4, IL‐6, IL‐8, IP10/CXCL10, LIF, MCP1, and M‐CSF) in the Control, Control + Ghrelin, D‐gal, and D‐gal + Ghrelin groups. *** p < 0.001.

    Article Snippet: Following antibody stripping and re‐blocking, sections were incubated overnight at 4°C with a second primary antibody: γH2AX (1:400; ab81299, Abcam), high‐mobility group box 1 (HMGB1) (1:400; AWA10032 , Abiowell), p21 Cip1 (1:200; ab188224, Abcam), p16 Ink4a (1:200; bs‐23797R, Bioss, China), or Lamin B1 (1:400; ab8982, Abcam).

    Techniques: Control, Western Blot, Analysis, Tissue, Immunofluorescence, Enzyme-linked Immunosorbent Assay

    Senescence was detected more frequently in MTJ-specific and vascular endothelial cells compared to the overall cell population. Representative in situ hybridisation images of young and old mouse MTJ (n = 4 per age group) showing senescence markers ( a ) p16 and ( b ) p21.Red: Col22 (MTJ marker); yellow: ( a ) p16 and ( b ) p21; cyan: VWF (endothelial cell marker); grey: DAPI. Blue DAPI image on the right shows the interface between the muscle and tendon indicated by orange dashed lines. Scale bar is 50 µm. Percentage of ( c ) p16 and ( d ) p21 positive cells were measured in Col22-positive MTJ-specific cells, VWF-positive endothelial cells, and all cells in the field of view. Y: young; O: old. Two-way ANOVA followed by Tukey’s multiple comparisons test was used to calculate the significance between the age groups and cell populations. Data are presented as mean ± SD: *p < 0.05 **p < 0.01 ****p < 0.0001

    Journal: Biogerontology

    Article Title: Ageing-related structural and cellular alterations in the mouse muscle-tendon junction

    doi: 10.1007/s10522-026-10428-x

    Figure Lengend Snippet: Senescence was detected more frequently in MTJ-specific and vascular endothelial cells compared to the overall cell population. Representative in situ hybridisation images of young and old mouse MTJ (n = 4 per age group) showing senescence markers ( a ) p16 and ( b ) p21.Red: Col22 (MTJ marker); yellow: ( a ) p16 and ( b ) p21; cyan: VWF (endothelial cell marker); grey: DAPI. Blue DAPI image on the right shows the interface between the muscle and tendon indicated by orange dashed lines. Scale bar is 50 µm. Percentage of ( c ) p16 and ( d ) p21 positive cells were measured in Col22-positive MTJ-specific cells, VWF-positive endothelial cells, and all cells in the field of view. Y: young; O: old. Two-way ANOVA followed by Tukey’s multiple comparisons test was used to calculate the significance between the age groups and cell populations. Data are presented as mean ± SD: *p < 0.05 **p < 0.01 ****p < 0.0001

    Article Snippet: The details of primary antibodies were as follows: Guinea‐pig anti‐collagen XXII (1:100, monoclonal, a kind gift from Manuel Koch, University of Cologne, Germany) and rabbit anti-p16 (1:50, 80772S, Cell Signaling Technology, MA, USA).

    Techniques: In Situ, Hybridization, Marker

    Protein expression analysis showed an increase in p16-positive cell proportion within Col22-positive MTJ-specific cells with age. a Representative immunofluorescent images of young and old MTJ (n = 4 per age group). Red: Col22; green: p16; grey: DAPI. Blue DAPI image on the right shows the interface between the muscle and tendon indicated by orange dashed lines. Scale bar is 50 µm. b Percentage of p16 positive cells in Col22-positive MTJ-specific cell population. Data are presented as mean ± SD (n = 4). A Mann-Whitney test was used to calculate the significance between young and old tendons. *p < 0.05

    Journal: Biogerontology

    Article Title: Ageing-related structural and cellular alterations in the mouse muscle-tendon junction

    doi: 10.1007/s10522-026-10428-x

    Figure Lengend Snippet: Protein expression analysis showed an increase in p16-positive cell proportion within Col22-positive MTJ-specific cells with age. a Representative immunofluorescent images of young and old MTJ (n = 4 per age group). Red: Col22; green: p16; grey: DAPI. Blue DAPI image on the right shows the interface between the muscle and tendon indicated by orange dashed lines. Scale bar is 50 µm. b Percentage of p16 positive cells in Col22-positive MTJ-specific cell population. Data are presented as mean ± SD (n = 4). A Mann-Whitney test was used to calculate the significance between young and old tendons. *p < 0.05

    Article Snippet: The details of primary antibodies were as follows: Guinea‐pig anti‐collagen XXII (1:100, monoclonal, a kind gift from Manuel Koch, University of Cologne, Germany) and rabbit anti-p16 (1:50, 80772S, Cell Signaling Technology, MA, USA).

    Techniques: Expressing, MANN-WHITNEY

    M6A modifications within total mRNA were significantly enhanced during the aging process . ( A ) Left: representative photographs of 2-month and 20-month-old male C57BL/6 mouse; Right: representative photographs of SA-β-gal staining in kidney tissues from young and old C57-BL/6 mice. ( B ) Left: representative photographs of SA-β-gal staining of P3 and P9 MEF cells; Right: the p16 expression levels in P3 and P9 MEF cells. ( C ) Left: representative photographs of SA-β-gal staining of H 2 O 2 -induced premature senescence of NIH/3T3 cells; Right: the p16 expression levels in H 2 O 2 treated NIH/3T3 cells. ( D ) Poly(A)+ RNA was extracted from the brain and liver tissue of young and old C57-BL/6 mice and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( E ) Poly(A)+ RNA was extracted from the P3 and P9 MEF cells and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( F ) Poly(A)+ RNA was extracted from the P3 and P9 MEF cells and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( G ) Expression levels of m6A modification associated proteins were measured in P3 and P9 MEF cells, and the quantitative results were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; One sample t test (MEF P9 vs 1, METTL3: p =0.0027, t = 9.176).

    Journal: Aging and Disease

    Article Title: The Flip Side of the Coin: METTL3 Serves as a Novel Cellular Senescence Accelerator via Negative Regulation of ITGA9

    doi: 10.14336/AD.2024.1715

    Figure Lengend Snippet: M6A modifications within total mRNA were significantly enhanced during the aging process . ( A ) Left: representative photographs of 2-month and 20-month-old male C57BL/6 mouse; Right: representative photographs of SA-β-gal staining in kidney tissues from young and old C57-BL/6 mice. ( B ) Left: representative photographs of SA-β-gal staining of P3 and P9 MEF cells; Right: the p16 expression levels in P3 and P9 MEF cells. ( C ) Left: representative photographs of SA-β-gal staining of H 2 O 2 -induced premature senescence of NIH/3T3 cells; Right: the p16 expression levels in H 2 O 2 treated NIH/3T3 cells. ( D ) Poly(A)+ RNA was extracted from the brain and liver tissue of young and old C57-BL/6 mice and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( E ) Poly(A)+ RNA was extracted from the P3 and P9 MEF cells and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( F ) Poly(A)+ RNA was extracted from the P3 and P9 MEF cells and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( G ) Expression levels of m6A modification associated proteins were measured in P3 and P9 MEF cells, and the quantitative results were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; One sample t test (MEF P9 vs 1, METTL3: p =0.0027, t = 9.176).

    Article Snippet: M6A antibody (202003, Synaptic Systems, Germany), GAPDH Monoclonal antibody (60004-1-Ig, Proteintech, China), METTL3 Monoclonal antibody (ab195352, Abcam, UK), FTO Polyclonal antibody (27226-1-AP, Proteintech, China), METTL14 Polyclonal antibody (AP22363a, Abgent, USA), ALKBH5 Polyclonal antibody (16837-1-AP, Proteintech, China), WTAP Monoclonal antibody (60188-1-Ig, Proteintech, China), p16 INK4A antibody (F-12) (sc-1661, Santa Cruz, USA), ITGA9 Polyclonal antibody (AF3827, R&D, USA), BrdU Monoclonal antibody (66241-1-Ig, Proteintech, China), Goat Anti-Mouse antibody (Alexa Fluor® 488) (SA00013-1, Proteintech, China), Goat anti-rabbit IgG (H+L) antibody (A0208, Beyotime, China), Goat anti-mouse IgG (H+L) antibody (A0216, Beyotime, China), Donkey anti-goat IgG (H+L) antibody (A0181, Beyotime, China).

    Techniques: Staining, Expressing, Dot Blot, Control, Modification

    METTL3 accelerated replicative senescence in MEFs . ( A ) The protein levels in MEFs transfected with siNC and siMETTL3-1/2/3. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (siMETTL3-1 vs 1: p =0.0113, t =9.328; siMETTL3-2VS 1: p =0.0022, t =21.05; siMETTL3-3 vs 1: p =0.0117, t = 9.155). ( B ) Poly(A)+ RNA was extracted from MEF cells transfected with siNC and siMETTL3 and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( C ) Western blot assay of p16 expression in MEFs transfected with siNC and siMETTL3. The quantitative results were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; One sample t test (siMETTL3 vs 1, p =0.0006, t =15.36). ( D ) Representative photographs of SA-β-gal staining of MEF cells transfected with siNC and siMETTL3 (×100). The quantitative results of SA-β-gal staining were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; Paired t test (siMETTL3 vs siNC, p =0.0025, t =9.407). ( E ) Representative photographs of cells stained with DAPI (blue fluorescence) and BrdU (green fluorescence) in siNC and siMETTL3 transfected MEF cells (scale bar: 400μm). The quantitative results of the ratio (BrdU stained cells / DAPI stained cells) were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; Paired t test (siMETTL3 vs siNC, p =0.009, t =6.065). ( F ) The mRNA levels in MEFs infected with Lenti-NC and METTL3 lentivirus. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (METTL3VS 1, p =0.0058, t =13.03). ( G ) Poly(A)+ RNA was extracted from MEF cells infected with Lenti-NC and METTL3 lentivirus and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( H ) Western blot assay of METTL3 and p16 expression in MEFs infected with METTL3 lentivirus. The quantitative results were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (METTL3 vs 1, p =0.0342, t =5.271). ( I ) Representative photographs of SA-β-gal staining of MEF cells infected with Lenti-NC and METTL3 lentivirus (×100). The quantitative results of SA-β-gal staining were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test (METTL3 vs Lenti-NC, p =0.0088, t =10.59). ( J ) Representative photographs of cells stained with DAPI (blue fluorescence) and BrdU (green fluorescence) in Lenti-NC and METTL3 lentivirus infected MEF cells (scale bar: 400μm). The quantitative results of the ratio (BrdU stained cells / DAPI stained cells) were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test (METTL3 vs Lenti-NC, p =0.0065, t =12.35).

    Journal: Aging and Disease

    Article Title: The Flip Side of the Coin: METTL3 Serves as a Novel Cellular Senescence Accelerator via Negative Regulation of ITGA9

    doi: 10.14336/AD.2024.1715

    Figure Lengend Snippet: METTL3 accelerated replicative senescence in MEFs . ( A ) The protein levels in MEFs transfected with siNC and siMETTL3-1/2/3. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (siMETTL3-1 vs 1: p =0.0113, t =9.328; siMETTL3-2VS 1: p =0.0022, t =21.05; siMETTL3-3 vs 1: p =0.0117, t = 9.155). ( B ) Poly(A)+ RNA was extracted from MEF cells transfected with siNC and siMETTL3 and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( C ) Western blot assay of p16 expression in MEFs transfected with siNC and siMETTL3. The quantitative results were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; One sample t test (siMETTL3 vs 1, p =0.0006, t =15.36). ( D ) Representative photographs of SA-β-gal staining of MEF cells transfected with siNC and siMETTL3 (×100). The quantitative results of SA-β-gal staining were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; Paired t test (siMETTL3 vs siNC, p =0.0025, t =9.407). ( E ) Representative photographs of cells stained with DAPI (blue fluorescence) and BrdU (green fluorescence) in siNC and siMETTL3 transfected MEF cells (scale bar: 400μm). The quantitative results of the ratio (BrdU stained cells / DAPI stained cells) were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; Paired t test (siMETTL3 vs siNC, p =0.009, t =6.065). ( F ) The mRNA levels in MEFs infected with Lenti-NC and METTL3 lentivirus. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (METTL3VS 1, p =0.0058, t =13.03). ( G ) Poly(A)+ RNA was extracted from MEF cells infected with Lenti-NC and METTL3 lentivirus and subjected to RNA dot-blot analysis with an antibody recognizing m6A. Methylene blue staining served as the loading control. ( H ) Western blot assay of METTL3 and p16 expression in MEFs infected with METTL3 lentivirus. The quantitative results were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (METTL3 vs 1, p =0.0342, t =5.271). ( I ) Representative photographs of SA-β-gal staining of MEF cells infected with Lenti-NC and METTL3 lentivirus (×100). The quantitative results of SA-β-gal staining were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test (METTL3 vs Lenti-NC, p =0.0088, t =10.59). ( J ) Representative photographs of cells stained with DAPI (blue fluorescence) and BrdU (green fluorescence) in Lenti-NC and METTL3 lentivirus infected MEF cells (scale bar: 400μm). The quantitative results of the ratio (BrdU stained cells / DAPI stained cells) were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test (METTL3 vs Lenti-NC, p =0.0065, t =12.35).

    Article Snippet: M6A antibody (202003, Synaptic Systems, Germany), GAPDH Monoclonal antibody (60004-1-Ig, Proteintech, China), METTL3 Monoclonal antibody (ab195352, Abcam, UK), FTO Polyclonal antibody (27226-1-AP, Proteintech, China), METTL14 Polyclonal antibody (AP22363a, Abgent, USA), ALKBH5 Polyclonal antibody (16837-1-AP, Proteintech, China), WTAP Monoclonal antibody (60188-1-Ig, Proteintech, China), p16 INK4A antibody (F-12) (sc-1661, Santa Cruz, USA), ITGA9 Polyclonal antibody (AF3827, R&D, USA), BrdU Monoclonal antibody (66241-1-Ig, Proteintech, China), Goat Anti-Mouse antibody (Alexa Fluor® 488) (SA00013-1, Proteintech, China), Goat anti-rabbit IgG (H+L) antibody (A0208, Beyotime, China), Goat anti-mouse IgG (H+L) antibody (A0216, Beyotime, China), Donkey anti-goat IgG (H+L) antibody (A0181, Beyotime, China).

    Techniques: Transfection, Dot Blot, Staining, Control, Western Blot, Expressing, Fluorescence, Infection

    ITGA9 staved off senescence in MEFs . ( A ) The mRNA and protein levels of ITGA9 in MEF cells transfected with siNC and siITGA9-1/2/3.Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (siITGA9-1 vs 11: p =0.0002, t =68.12; siITGA9-2 vs 1: p =0.0003, t =63.15 ; siITGA9-3 vs 11: p =0.0111, t =9.410; ) (B) The p16 expression levels in MEF cells transfected with siNC and siITGA9. The quantitative results were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (siITGA9 vs 1, p =0.0012, t =28.41). ( C ) Representative photographs of SA-β-gal staining of MEF cells transfected with siNC and siITGA9 (×100). The quantitative results of SA-β-gal staining were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test (siITGA9 vs siNC, p =0.0026, t =19.65). ( D ) Representative photographs of cells stained with DAPI (blue fluorescence) and BrdU (green fluorescence) in siNC and siITGA9 transfected MEF cells (scale bar: 400μm). The quantitative results of the ratio ( BrdU stained cells / DAPI stained cells) were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test (siITGA9 vs siNC, p =0.0144, t =4.138). ( E ) Representative photographs of cells stained with DAPI (blue fluorescence) and BrdU (green fluorescence) in Lenti-NC and ITGA9 infected MEF cells (scale bar: 400μm). The quantitative results of the ratio ( BrdU stained cells / DAPI stained cells) were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; Paired t test (ITGA9 vs Lenti-NC, p =0.01, t =5.843). ( F ) The mRNA levels of ITGA9 in MEF cells infected with Lenti-NC or ITGA9 lentivirus. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (ITGA9 vs 1, p =0.0025, t =20.08). ( G ) The ITGA9 and p16 protein expression levels in MEFs infected with Lenti-NC and ITGA9 lentivirus. The results of grayscale scanning were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (ITGA9 vs 1, p =0.0256, t =6.124). ( H ) Representative photographs of SA-β-gal staining of MEF cells infected with Lenti-NC and ITGA9 lentivirus (×100). The quantitative results of SA-β-gal staining were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test (ITGA9 vs Lenti-NC, p =0.0009, t =33.30).

    Journal: Aging and Disease

    Article Title: The Flip Side of the Coin: METTL3 Serves as a Novel Cellular Senescence Accelerator via Negative Regulation of ITGA9

    doi: 10.14336/AD.2024.1715

    Figure Lengend Snippet: ITGA9 staved off senescence in MEFs . ( A ) The mRNA and protein levels of ITGA9 in MEF cells transfected with siNC and siITGA9-1/2/3.Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (siITGA9-1 vs 11: p =0.0002, t =68.12; siITGA9-2 vs 1: p =0.0003, t =63.15 ; siITGA9-3 vs 11: p =0.0111, t =9.410; ) (B) The p16 expression levels in MEF cells transfected with siNC and siITGA9. The quantitative results were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (siITGA9 vs 1, p =0.0012, t =28.41). ( C ) Representative photographs of SA-β-gal staining of MEF cells transfected with siNC and siITGA9 (×100). The quantitative results of SA-β-gal staining were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test (siITGA9 vs siNC, p =0.0026, t =19.65). ( D ) Representative photographs of cells stained with DAPI (blue fluorescence) and BrdU (green fluorescence) in siNC and siITGA9 transfected MEF cells (scale bar: 400μm). The quantitative results of the ratio ( BrdU stained cells / DAPI stained cells) were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test (siITGA9 vs siNC, p =0.0144, t =4.138). ( E ) Representative photographs of cells stained with DAPI (blue fluorescence) and BrdU (green fluorescence) in Lenti-NC and ITGA9 infected MEF cells (scale bar: 400μm). The quantitative results of the ratio ( BrdU stained cells / DAPI stained cells) were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; Paired t test (ITGA9 vs Lenti-NC, p =0.01, t =5.843). ( F ) The mRNA levels of ITGA9 in MEF cells infected with Lenti-NC or ITGA9 lentivirus. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (ITGA9 vs 1, p =0.0025, t =20.08). ( G ) The ITGA9 and p16 protein expression levels in MEFs infected with Lenti-NC and ITGA9 lentivirus. The results of grayscale scanning were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; One sample t test (ITGA9 vs 1, p =0.0256, t =6.124). ( H ) Representative photographs of SA-β-gal staining of MEF cells infected with Lenti-NC and ITGA9 lentivirus (×100). The quantitative results of SA-β-gal staining were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test (ITGA9 vs Lenti-NC, p =0.0009, t =33.30).

    Article Snippet: M6A antibody (202003, Synaptic Systems, Germany), GAPDH Monoclonal antibody (60004-1-Ig, Proteintech, China), METTL3 Monoclonal antibody (ab195352, Abcam, UK), FTO Polyclonal antibody (27226-1-AP, Proteintech, China), METTL14 Polyclonal antibody (AP22363a, Abgent, USA), ALKBH5 Polyclonal antibody (16837-1-AP, Proteintech, China), WTAP Monoclonal antibody (60188-1-Ig, Proteintech, China), p16 INK4A antibody (F-12) (sc-1661, Santa Cruz, USA), ITGA9 Polyclonal antibody (AF3827, R&D, USA), BrdU Monoclonal antibody (66241-1-Ig, Proteintech, China), Goat Anti-Mouse antibody (Alexa Fluor® 488) (SA00013-1, Proteintech, China), Goat anti-rabbit IgG (H+L) antibody (A0208, Beyotime, China), Goat anti-mouse IgG (H+L) antibody (A0216, Beyotime, China), Donkey anti-goat IgG (H+L) antibody (A0181, Beyotime, China).

    Techniques: Transfection, Expressing, Staining, Fluorescence, Infection

    METTL3 accelerated cellular senescence by modulating ITGA9 . ( A ) The p16 level in MEF cells transfected with siNC, siMETTL3 or siMETTL3 and siITGA9. The quantitative results were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; Paired t test ( p =0.0058, t =7.062). ( B ) The p16 level in MEF cells infected with Lenti-NC, METTL3 or METTL3 and ITGA9. The quantitative results were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test ( p =0.017, t =7.563). (C, D) Representative photographs of SA-β-gal staining (C) and BrdU incorporation staining (D) of MEF cells transfected with siNC, siMETTL3, siITGA9, or siMETTL3 and siITGA9. The quantitative results were shown on the right. Mean (±SEM), passed normality test: Shapiro-Wilk test; Paired t test (C), n=3, p =0.0044, t =15.02; (D), n=4, p =0.0003, t =19.14). (E, F) Representative photographs of SA-β-gal staining (E) and BrdU incorporation staining (F) of MEF cells infected with Lenti-NC, METTL3, ITGA9 or METTL3 and ITGA9. The quantitative results were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test ((E): p =0.0095, t =10.18; (F): p =0.0231, t =6.469).

    Journal: Aging and Disease

    Article Title: The Flip Side of the Coin: METTL3 Serves as a Novel Cellular Senescence Accelerator via Negative Regulation of ITGA9

    doi: 10.14336/AD.2024.1715

    Figure Lengend Snippet: METTL3 accelerated cellular senescence by modulating ITGA9 . ( A ) The p16 level in MEF cells transfected with siNC, siMETTL3 or siMETTL3 and siITGA9. The quantitative results were shown on the right. Mean (±SEM), n=4, passed normality test: Shapiro-Wilk test; Paired t test ( p =0.0058, t =7.062). ( B ) The p16 level in MEF cells infected with Lenti-NC, METTL3 or METTL3 and ITGA9. The quantitative results were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test ( p =0.017, t =7.563). (C, D) Representative photographs of SA-β-gal staining (C) and BrdU incorporation staining (D) of MEF cells transfected with siNC, siMETTL3, siITGA9, or siMETTL3 and siITGA9. The quantitative results were shown on the right. Mean (±SEM), passed normality test: Shapiro-Wilk test; Paired t test (C), n=3, p =0.0044, t =15.02; (D), n=4, p =0.0003, t =19.14). (E, F) Representative photographs of SA-β-gal staining (E) and BrdU incorporation staining (F) of MEF cells infected with Lenti-NC, METTL3, ITGA9 or METTL3 and ITGA9. The quantitative results were shown on the right. Mean (±SEM), n=3, passed normality test: Shapiro-Wilk test; Paired t test ((E): p =0.0095, t =10.18; (F): p =0.0231, t =6.469).

    Article Snippet: M6A antibody (202003, Synaptic Systems, Germany), GAPDH Monoclonal antibody (60004-1-Ig, Proteintech, China), METTL3 Monoclonal antibody (ab195352, Abcam, UK), FTO Polyclonal antibody (27226-1-AP, Proteintech, China), METTL14 Polyclonal antibody (AP22363a, Abgent, USA), ALKBH5 Polyclonal antibody (16837-1-AP, Proteintech, China), WTAP Monoclonal antibody (60188-1-Ig, Proteintech, China), p16 INK4A antibody (F-12) (sc-1661, Santa Cruz, USA), ITGA9 Polyclonal antibody (AF3827, R&D, USA), BrdU Monoclonal antibody (66241-1-Ig, Proteintech, China), Goat Anti-Mouse antibody (Alexa Fluor® 488) (SA00013-1, Proteintech, China), Goat anti-rabbit IgG (H+L) antibody (A0208, Beyotime, China), Goat anti-mouse IgG (H+L) antibody (A0216, Beyotime, China), Donkey anti-goat IgG (H+L) antibody (A0181, Beyotime, China).

    Techniques: Transfection, Infection, Staining, BrdU Incorporation Assay