tunel staining kit  (Servicebio Inc)

Journal: Investigative Ophthalmology & Visual Science

Article Title: YBX1 Modulated Corneal Neovascularization Induced by Alkali Burn via m 5 C-Dependent Regulation of the STAT3/HIF-1α/VEGFA Axis

doi: 10.1167/iovs.67.2.42

Figure Lengend Snippet: YBX1 regulates angiogenesis and apoptosis in HUVECs through the HIF-1α pathway. ( A–H ) Western blot results and quantification of HIF-1α, VEGFA, Bcl-2, Bax, cleaved PARP, and cleaved Caspase-3 expression levels after overexpression of HIF-1α in the context of YBX1 knockdown. ( I ) TUNEL staining of control, H/R and different treatment groups evaluating by fluorescence microscope. Magnification = 40×; Scale bar = 50 µm. ( J ) Double ICC labeling for VEGFA and CD31 in HUVECs with YBX1 knockdown and HIF-1α overexpression. Magnification = 400×; Scale bar = 50 µm. ( K ) Scratch wound healing assay showing changes in cell migration ability among different treatment groups. Magnification = 100×; Scale bar = 500 µm. ( L ) Transwell migration assay assessing invasion capacity after YBX1 knockdown and HIF-1α overexpression. Magnification = 400×; Scale bar = 125 µm. * P < 0.05 versus the control group, # P < 0.05 versus the H/R group, ◆ P < 0.05 versus the H/R + siNC group, & P < 0.05 versus the H/R + siYBX1. Data are presented as mean ± SEM ( n = 6). Means were compared by 1-way ANOVA with Tukey's multiple comparison post hoc tests.

Article Snippet: Cell apoptosis was assessed via TUNEL staining (Vazyme, Cat #A112).

Techniques: Western Blot, Expressing, Over Expression, Knockdown, TUNEL Assay, Staining, Control, Fluorescence, Microscopy, Labeling, Wound Healing Assay, Migration, Transwell Migration Assay, Comparison

Journal: Bioactive Materials

Article Title: PKM2 + apoptotic vesicle-mediated systemic senolytics ameliorate chronic periodontitis

doi: 10.1016/j.bioactmat.2025.06.041

Figure Lengend Snippet: Irreversible periodontitis leads to apoptotic resistance. ( a – c ) Western blotting was used to evaluate the Fas, Caspase 3, Cleaved caspase 3 (Cl-caspase 3), Bax, and Bcl-xl expression in the mouse spleen, bone marrow, and peripheral blood lymphocytes. ( d ) The expression of Cl-caspase 3 and Bax human peripheral blood lymphocytes was evaluated by Western blotting. ( e and f ) Mouse peripheral blood lymphocyte apoptotic rates, as determined by flow cytometry. (n = 4) ( g and h ) Human peripheral blood lymphocyte apoptotic rate was analyzed by flow cytometry. (n = 4) ( i and j ) IF staining and quantification of TUNEL in the spleen and femur. (n = 4) Scale bar, 50 μm. ( k ) The p65, p-p65, p16, and Bcl-xl expression of BMMSCs was evaluated by Western blotting. ( l and m ) SA-β-Gal staining was used to analyze senescence of BMMSCs. (n = 4) Scale bar, 50 μm. ( n and o ) Flow cytometry was used to determine the apoptotic rate of BMMSCs. (n = 4) For two-group comparisons, significance was assessed by unpaired Student's t -test. Error bars show the mean ± SD. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. ns, not significant.

Article Snippet: Apoptotic cells were detected using a TUNEL staining kit (Servicebio, China).

Techniques: Western Blot, Expressing, Flow Cytometry, Staining, TUNEL Assay

Journal: Bioactive Materials

Article Title: PKM2 + apoptotic vesicle-mediated systemic senolytics ameliorate chronic periodontitis

doi: 10.1016/j.bioactmat.2025.06.041

Figure Lengend Snippet: The senolytics dasatinib and quercetin can reverse alveolar bone loss in CP mice. ( a ) Representative microCT 3-dimensional reconstructed images of maxillae and HE staining on maxilla sections in CP mice. Ctrl: Normal mice without any treatment, used as baseline controls, Veh: CP mice treated with vehicle solution (10 % polyethylene glycol 4000), D + Q: CP mice treated with dasatinib (5 mg/kg body mass) and quercetin (50 mg/kg body mass), Scale bar, 0.4 μm (microCT), 200 μm (HE). ( b ) Quantification of alveolar bone loss, represented by measurement of palatal CEJ-ABC area (red line surrounding area). (n = 4–5) ( c and d ) TRAP staining showed a reduction in osteoclasts (stained in red) in the D + Q group compared to the vesicle control (Veh) group. (n = 3) Scale bar, 20 μm. ( e ) ELISA showed proinflammatory cytokines IL-17, IL-1β, TNF-α, TNF-β, IFN-γ, and IL-6 in the plasma. D + Q significantly reduced TNF-β and IFN-γ levels in periodontitis mice. (n = 4) ( f – i ) IF staining of p16 and SA-β-Gal staining revealed that D + Q reduced the accumulation of senescent cells in the liver, spleen, kidney, and femur. (n = 4) Scale bar, 10 μm (IF), 50 μm (β-Gal). ( j and k ) IF staining and quantification of TUNEL in the spleen and femur. (n = 4) Scale bar, 50 μm. Differences among three groups were analyzed by one-way ANOVA with Tukey's post hoc test. Error bars show the mean ± SD. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. ns, not significant.

Article Snippet: Apoptotic cells were detected using a TUNEL staining kit (Servicebio, China).

Techniques: Staining, Control, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, TUNEL Assay

Journal: Bioactive Materials

Article Title: PKM2 + apoptotic vesicle-mediated systemic senolytics ameliorate chronic periodontitis

doi: 10.1016/j.bioactmat.2025.06.041

Figure Lengend Snippet: MSC-apoV and MSC infusion ameliorates local alveolar bone loss and systemic inflammation in periodontitis mice. ( a and b ) Representative microCT 3-dimensional reconstructed images of maxillae and HE staining on maxilla sections showed that both MSC-apoV and MSC infusion could promote alveolar bone repair. The data points represent measurements from both the left and right sides of each mouse. Due to unsuccessful sample collection, some mice contributed data from only one side, which creates a sample size that seems to be less than 4 in the illustration. This has been clarified to avoid confusion. PBS: CP mice treated with PBS, MSCs:CP mice treated with MSCs, ApoVs: CP mice treated with MSC-apoVs, (n = 4–5) Scale bar, 0.4 mm (microCT), 1000 μm (HE). ( c and d ) TRAP staining showed more osteoclasts (stained in red) in the PBS group compared to the MSC-apoV and MSC groups. (n = 3) Scale bar, 20 μm. ( e ) HE staining demonstrated that MSC-apoVs and MSCs rescued pathological changes in the liver, spleen, kidney, and femur. Scale bar, 50 μm. ( f – i ) MSC-apoVs and MSCs decreased senescent cells in the liver, spleen, kidney, and femur, which were analyzed by SA-β-Gal staining and p16 IF staining. (n = 4) Scale bar, 50 μm (β-Gal), 10 μm (IF). ( j and k ) TUNEL staining was used to evaluate the apoptotic levels in the spleen and bone marrow. Both MSC-apoVs and MSCs promoted apoptosis in these tissues. (n = 4) Differences among three groups were analyzed by one-way ANOVA with Tukey's post hoc test. Scale bar, 50 μm. Error bars show the mean ± SD. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. ns, not significant.

Article Snippet: Apoptotic cells were detected using a TUNEL staining kit (Servicebio, China).

Techniques: Staining, TUNEL Assay

Journal: Bioactive Materials

Article Title: PKM2 + apoptotic vesicle-mediated systemic senolytics ameliorate chronic periodontitis

doi: 10.1016/j.bioactmat.2025.06.041

Figure Lengend Snippet: PKM2 is responsible for MSC-apoV-mediated amelioration of local alveolar bone loss and systemic inflammation in periodontitis mice. ( a ) Representative microCT 3-dimensional reconstructed images of maxillae and HE staining on maxilla sections showed that PKM2 + MSC-apoV infusion could promote alveolar bone repair, while PKM2 low MSC-apoVs failed to do so. PBS: CP mice treated with PBS, PKM2 + :CP mice treated with PKM2 + MSC-apoVs, PKM low : CP mice treated with PKM2 low MSC-apoVs, Scale bar, 0.4 mm (microCT), 1000 μm (HE). ( b ) Quantification of periodontitis-induced alveolar bone loss, represented by measurement of palatal CEJ-ABC area (red line surrounding area). (n = 4–5) ( c and d ) Representative images and quantification of TRAP staining showed more osteoclasts (stained in red) in the PBS and PKM2 low MSC-apoV groups compared to the PKM2 + MSC-apoV group. (n = 3) Scale bar, 20 μm. ( e ) PKM2 + MSC-apoVs alleviated proinflammatory cytokines TNF-α, TNF-β, and IFN-γ. Cytokines in the plasma were analyzed using ELISA. (n = 3–4) ( f ) Flow cytometry demonstrated the ratios of Th17/Treg were decreased in peripheral blood, after treatment with PKM2 + MSC-apoVs. (n = 4–5) ( g ) HE staining demonstrated that PKM2 + MSC-apoVs alleviated pathological changes in the liver, spleen, kidneys, and femur whereas PBS and PKM2 low MSC-apoV groups did not. Scale bar, 50 μm. ( h – k ) SA-β-Gal staining and p16 IF staining revealed that PKM2 + MSC-apoVs, but not PKM2 low MSC-apoVs, reduced the accumulation of senescent cells in the liver, spleen, kidney, and femur. (n = 4) Scale bar, 50 μm (β-Gal), 200 μm (β-Gal spleen), 10 μm (IF). ( l and m ) Apoptotic levels of spleen and bone marrow were evaluated by TUNEL staining. PKM2 + MSC-apoVs increased apoptosis of spleen and bone marrow. (n = 4–5) Scale bar, 50 μm. Differences among three groups were analyzed by one-way ANOVA with Tukey's post hoc test. Error bars show the mean ± SD. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. ns, not significant.

Article Snippet: Apoptotic cells were detected using a TUNEL staining kit (Servicebio, China).

Techniques: Staining, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Flow Cytometry, TUNEL Assay

Journal: bioRxiv

Article Title: Curcumin activates distinct programming of redox metabolism towards differential regulation of gene expression mediated by Nrf1 and Nrf2

doi: 10.1101/2025.07.09.663854

Figure Lengend Snippet: (A) The in vivo malgrowth curve (a1) of xenograft tumors in distinct groups of nude mice, that were subcutaneously injected with human WT or Nrf1α –/– hepatoma cell lines, followed by intervention with CUR (I.G., 35 mg/mL) or vehicle (I.G., 0.9% physiological saline). And the volumes (a2) and weight (a3) of the above xenograft tumors were determined as shown graphically. (B) The model tumor tissues were subjected to the histopathological examination by routine HE staining, and then relevant images were obtained from microscopy. (C) The staining of TUNEL was subjected to evaluating cell apoptosis in xenograft tumor tissues. (D-F) The content of glycose (D) , the ratio of ATP (E) and NADP + /NADPH (F) within tumor tissues were detected, which were performed at least in triplicates. The obtained absorbance (OD) values are statistically determined and shown graphically, with significant increases ($, p <0.05) and significant decreases ( *p < 0.05), relative to the control value of each group as indicated. Additional symbols “$ or *” indicate significant differences in tumor tissues compared to their respective t0 value measured after CUR intervention. (G) Schematic diagram shows tryptophan metabolic pathway (adapted from the online KEGG pathway database). The “ black ” means metabolic compounds, “ red ” represents key metabolites, and “ blue ” represents different metabolic enzymes. (H) CUR differential expression heat map of related metabolites in tryptophan metabolism in different tumor tissues (i.e., l Log 2 FC l≥1). (L) Real-time qPCR analysis was subjected to detecting the effects of CUR treatment on transcriptional expression of those key enzymes (e.g., Nrf1, TDO, KMO, CYP1A1, QPRT, Nrf2, HAAO, DDC, AhR, AFMID, KYN, AOX1, ASMT, AADAT, AANAT, TPH2 ) in tryptophan metabolism pathway. The resulting data were shown as fold changes (mean±SD, n =3×3), which are representative of at least three independent experiments being each performed in triplicates. Significant increases ($, p < 0.05; $$, p < 0.01) and significant decreases ( *p < 0.05), were statistically analyzed when compared to WT_vehicle , respectively. Additional symbols “$ or *” indicate significant differences in Nrf1α –/– tumors as compared to their vehicle controls. “ND” means no significant different. (M) Western blotting analysis was subjected to detecting the protein expression of Nrf2, IDO2, MAOB and KYNU in different tumor tissues. The intensity of those immunoblots was also quantified by the Quantity One 4.5.2 software, and showed below the indicated protein bands. (K) Overview of the effects of CUR on different mechanisms through Nrf1/2 differential expression at the cellular and animal levels.

Article Snippet: The subsequent experimental procedure was manipulated according to the instruction of an in situ Tunel (TdT-mediated dUTP Nick-End Labeling) fluorescent staining kit (G002-3-1, Nanjing Jiancheng, Nanjing, China), before being stained with DAPI.

Techniques: In Vivo, Injection, Saline, Staining, Microscopy, TUNEL Assay, Control, Quantitative Proteomics, Expressing, Western Blot, Software