Journal: Biology Direct

Article Title: V-ATPase-targeted siRNA library screening reveals ATP6V1A negatively regulates UVB-induced keratinocyte senescence

doi: 10.1186/s13062-026-00891-4

Figure Lengend Snippet: Lysosomes are damaged in UVB-induced senescent cells. ( A ) Representative images of SA-β-gal staining in HaCaT cells after UVB-induced senescence. Scale bar = 25 μm. Right panel showed the mean SA-β-gal positivity rate ( n = 5). ( B ) Changes in LysoSensor staining in HaCaT cells after UVB-induced senescence. Scale bar = 20 μm. Right is the quantitative results of LysoSensor fluorescence intensity. ( C ) Western blot and grayscale analysis of CTSD in HaCaT cells after UVB-induced senescence. Grayscale normalized to β-actin of sham cells. ( D ) IF detection of Galectin-3 and LAMP1 expression levels in HaCaT cells after UVB-induced senescence. Scale bar = 5 μm. E , F . The colocalization analysis and line-scan intensity profiles of Galectin-3 and LAMP1 of Fig. D. ( G ) Quantitative results for the fluorescence intensity of Galectin-3 and LAMP1 in Fig. D. ( H ) GO analysis of aging and young skin in GSE130973 . Higher GeneRatio indicates greater pathway relevance. ( I ) Correlation analysis of V-ATPase complex expression and aging score in keratinocytes. Data are expressed as mean ± standard deviation (SD). Student’s t-test. ns indicates no statistical significance. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001

Article Snippet: SA-β-gal staining was used the SA-β-gal assay kit (Beyotime, China).

Techniques: Staining, Fluorescence, Western Blot, Analysis, Expressing, Standard Deviation

Journal: Biology Direct

Article Title: V-ATPase-targeted siRNA library screening reveals ATP6V1A negatively regulates UVB-induced keratinocyte senescence

doi: 10.1186/s13062-026-00891-4

Figure Lengend Snippet: ATP6V1A expression decreases following UVB-induced senescence. ( A ) Schematic of siRNA libraries screening combined with SPiDER-βgal detection and the heat map of the results. ( B ) Z -score for the 7 genes with the most significant upregulation in screening. ( C ) IF was used to observe the expression of ATP6V1A and LAMP1 in UVB-induced senescent HaCaT cells. Scale bar = 10 μm. Right are the quantitative results for the fluorescence intensity of ATP6V1A and LAMP1. ( D ) IF analysis of p16 expression in the dorsal skin of mice following UVB-induced senescence. Scale bar = 12.5 μm. Right is the quantitative results of p16 fluorescence intensity. ( E ) Verification of the UVB-induced senescent mice epidermis using SA-β-gal. Scale bar = 50 μm. The right panel shows the mean SA-β-gal positivity rate ( n = 5). ( F ) IF analysis of ATP6V1A and LAMP1 expression in the epidermis and dermis of the dorsal skin of UVB-induced senescent mice. Scale bar = 25 μm. G , H . The quantitative results for the fluorescence intensity of ATP6V1A in the epidermis and dermis of Fig. F. I , J . The quantitative results for the fluorescence intensity of LAMP1 in the epidermis and dermis of Fig. F. Data are expressed as mean ± standard deviation (SD). Student’s t-test. ns indicates no statistical significance. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001

Article Snippet: SA-β-gal staining was used the SA-β-gal assay kit (Beyotime, China).

Techniques: Expressing, Screening, Fluorescence, Analysis, Standard Deviation

Journal: Biology Direct

Article Title: V-ATPase-targeted siRNA library screening reveals ATP6V1A negatively regulates UVB-induced keratinocyte senescence

doi: 10.1186/s13062-026-00891-4

Figure Lengend Snippet: ATP6V1A regulates the UVB-induced senescence process in keratinocytes. ( A ) Western blot and grayscale analysis were used to observe the effects of ATP6V1A knockdown in HaCaT cells. Grayscale normalized to β-actin of siNC cells. ( B ) Representative SA-β-gal staining images of HaCaT cells following UVB-induced senescence after ATP6V1A knockdown. Scale bar = 25 μm. The right panel shows the mean SA-β-gal positivity rate ( n = 5). ( C ) Representative images of p16 and γH2A.X expression in HaCaT cells undergoing UVB-induced senescence following ATP6V1A knockdown. Scale bar = 10 μm. Below are the quantification results for p16 fluorescence and γH2A.X foci. ( D ) Western Blot and grayscale analysis were used to observe the effects of ATP6V1A overexpression with HA tag in HaCaT cells. Grayscale normalized to β-actin of NC cells. ( E ) Representative SA-β-gal staining images of HaCaT cells following UVB-induced senescence after ATP6V1A overexpression. Scale bar = 25 μm. Right panel showed the mean SA-β-gal positivity rate ( n = 5). ( F ) Representative images of p16 and γH2A.X expression in HaCaT cells undergoing UVB-induced senescence following ATP6V1A overexpression. Scale bar = 10 μm. Below are the quantification results for p16 fluorescence and γH2A.X foci. Data are expressed as mean ± standard deviation (SD). two-way ANOVA analysis. ns indicates no statistical significance. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001

Article Snippet: SA-β-gal staining was used the SA-β-gal assay kit (Beyotime, China).

Techniques: Western Blot, Analysis, Knockdown, Staining, Expressing, Fluorescence, Over Expression, Standard Deviation

Journal: Biomedicines

Article Title: Isoproterenol Induces Cardiac Injury and Senescence in Sprague–Dawley Rats: A Cost-Effective Pharmacological Model

doi: 10.3390/biomedicines14071445

Figure Lengend Snippet: ISO induces senescence in the apex of the young adult rat. 10–12-week-old rats were administered 100 mg/kg ISO or saline for five days, and then apexes were collected on days 10 and 28. ( A ) Representative SA-β-gal staining images (blue; black arrows). ( B ) Quantification of SA-β-gal-positive area. ( C ) Representative p16 staining images (green; yellow arrows). ( D ) Quantification of p16-positive cells. ( E ) Representative p21 staining images (green; yellow arrows). ( F ) Quantification of p21-positive cells. Scale bars = 100 µm. Data are expressed as the mean ± S.D. Statistical analysis was performed using two-way ANOVA followed by Šídák’s multiple comparisons test. n = 5 per group.

Article Snippet: Frozen sections were used for SA-β-gal (HY-K1089, MedChemExpress, Monmouth Junction, NJ, USA) and p16 staining.

Techniques: Saline, Staining

Journal: Biochemistry and Biophysics Reports

Article Title: Screening of metabolic-related biomarkers linking intervertebral disc degeneration and type 2 diabetes based on comprehensive bioinformatics analysis and machine learning

doi: 10.1016/j.bbrep.2026.102593

Figure Lengend Snippet: BCAA metabolism and immune changes in degenerated NP cells. (A) qRT-PCR was conducted to analysis the mRNA levels of BCAA metabolism-related enzymes and BCAA transporter in grade I/II and grade III/IV NP tissues; n = 20; P < 0.05. (B, C) Expression levels of mRNA for pro-inflammatory factors, BCAA metabolism-related enzymes, and BCAA transporters in control group and TNF-α-treated HNPC cells. (D) Expression of SA-β-gal in HNPC cells stimulated with different concentrations of TNF-α. (E, F) p16, p21, p53 mRNA level in TNF-α-stimulated HNPC cells,and correlation analysis with TNF-α concentration.

Article Snippet: Cellular senescence was assessed using Senescence-Associated β-Galactosidase (SA-β-gal) staining (Senescence β-Galactosidase Staining Kit, #C0602, Beyotime, China), performed according to the manufacturer's protocol.

Techniques: Quantitative RT-PCR, Expressing, Control, Concentration Assay