Journal: Biology

Article Title: Rapalink-1 Attenuates Oxidative-Stress-Induced Senescence in Vascular Cells in Association with Reduced NF-κB and MAPK Signaling

doi: 10.3390/biology15090732

Figure Lengend Snippet: ( A ) HUVECs were treated with increasing concentrations of H 2 O 2 (50–300 µM) for 24 h, and cell viability was assessed by MTT assay. ( B ) HUVECs were exposed to 300 µM H 2 O 2 in the presence of increasing concentrations of Rapalink-1 (50–400 pM) for 24 h, followed by viability measurement. Data are presented as mean ± SEM ( n = 3 independent experiments). Statistical significance was determined by one-way ANOVA with appropriate post hoc tests. Significance is indicated as: * p < 0.05, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human umbilical-vein endothelial-cell (HUVEC) models were purchased from PromoCell (Heidelberg, Germany).

Techniques: MTT Assay

Journal: Biology

Article Title: Rapalink-1 Attenuates Oxidative-Stress-Induced Senescence in Vascular Cells in Association with Reduced NF-κB and MAPK Signaling

doi: 10.3390/biology15090732

Figure Lengend Snippet: ( A ) MTT assay results in HUVECs. ( B , C ) Representative images of DCFH-DA staining (green), Hoechst counterstaining (blue), merged images and quantification of DCFH-DA fluorescence intensity in HUVECs. ( D – F ) qPCR analysis in HUVECs: NRF2 , NOX4 , MnSOD . Scale bar = 50 μm. Data are presented as mean ± SD ( n = 3). Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test. Significance is indicated as: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human umbilical-vein endothelial-cell (HUVEC) models were purchased from PromoCell (Heidelberg, Germany).

Techniques: MTT Assay, Staining, Fluorescence

Journal: Biology

Article Title: Rapalink-1 Attenuates Oxidative-Stress-Induced Senescence in Vascular Cells in Association with Reduced NF-κB and MAPK Signaling

doi: 10.3390/biology15090732

Figure Lengend Snippet: ( A , B ) Representative immunofluorescence images and quantification of γ-H2AX (green) in HUVECs with Hoechst nuclear counterstain (blue) and merged images. ( C , D ) Representative images and quantification of 8-OHDG (red) in HUVECs with Hoechst nuclear counterstain (blue) and merged images. Scale bar = 50 μm. Data are presented as mean ± SD ( n = 3). Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test. Significance is indicated as: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human umbilical-vein endothelial-cell (HUVEC) models were purchased from PromoCell (Heidelberg, Germany).

Techniques: Immunofluorescence

Journal: Biology

Article Title: Rapalink-1 Attenuates Oxidative-Stress-Induced Senescence in Vascular Cells in Association with Reduced NF-κB and MAPK Signaling

doi: 10.3390/biology15090732

Figure Lengend Snippet: ( A , B ) Representative images and quantification of SA-β-gal staining in HUVECs. ( C , D ) Representative immunofluorescence images and quantification of Lamin B1 (red) in HUVECs with Hoechst counterstain (blue) and merged images. ( E , F ) Representative Western blots and quantification of the senescence-associated cell cycle regulator p21 in HUVECs. ( G ) qPCR analysis of p16 mRNA expression in HUVECs. Protein expression was normalized to GAPDH. Scale bar = 50 μm. Data are presented as mean ± SD ( n = 3). Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test. Significance is indicated as: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human umbilical-vein endothelial-cell (HUVEC) models were purchased from PromoCell (Heidelberg, Germany).

Techniques: Staining, Immunofluorescence, Western Blot, Expressing

Journal: Biology

Article Title: Rapalink-1 Attenuates Oxidative-Stress-Induced Senescence in Vascular Cells in Association with Reduced NF-κB and MAPK Signaling

doi: 10.3390/biology15090732

Figure Lengend Snippet: ( A – F ) qPCR analysis of SASP-related inflammatory, cytokine, adhesion-related, and matrix-remodeling factors in HUVECs, including COX2 , IL-6 , IL-8 , ICAM1 , MMP1 , and TIMP1 . ( G – I ) Representative Western blots and quantification of relative protein expression of MMP2 and VCAM1 in HUVECs. Protein expression was normalized to GAPDH. Data are presented as mean ± SD ( n = 3). Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test. Significance is indicated as: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human umbilical-vein endothelial-cell (HUVEC) models were purchased from PromoCell (Heidelberg, Germany).

Techniques: Western Blot, Expressing

Journal: Biology

Article Title: Rapalink-1 Attenuates Oxidative-Stress-Induced Senescence in Vascular Cells in Association with Reduced NF-κB and MAPK Signaling

doi: 10.3390/biology15090732

Figure Lengend Snippet: ( A ) Representative Western blot images showing the protein levels of p65, p-p65, p38, p-p38, ERK, and p-ERK (HUVECs). ( B – D ) Quantitative analysis of p-p65/p65, p-p38/p38, and p-ERK/ERK (HUVECs). Protein expression was normalized to GAPDH. Data are presented as mean ± SD ( n = 3). Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test. Significance is indicated as: ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human umbilical-vein endothelial-cell (HUVEC) models were purchased from PromoCell (Heidelberg, Germany).

Techniques: Western Blot, Expressing

Journal: Biology

Article Title: Rapalink-1 Attenuates Oxidative-Stress-Induced Senescence in Vascular Cells in Association with Reduced NF-κB and MAPK Signaling

doi: 10.3390/biology15090732

Figure Lengend Snippet: ( A ) Representative Western blot images showing the protein levels of mTOR, p-mTOR, p-S6, p-4EBP1 (HUVECs). ( B – E ) Quantitative analysis of p-mTOR/mTOR, p-S6, p-4EBP1, p-AKT (HUVECs). Protein expression was normalized to GAPDH. Data are presented as mean ± SD ( n = 3). Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test. Significance is indicated as: ** p < 0.01, *** p < 0.001.

Article Snippet: Human umbilical-vein endothelial-cell (HUVEC) models were purchased from PromoCell (Heidelberg, Germany).

Techniques: Western Blot, Expressing