Journal: Cells

Article Title: Dysregulation of Caveolin-1 Phosphorylation and Nuclear Translocation Is Associated with Senescence Onset

doi: 10.3390/cells10112939

Figure Lengend Snippet: Cav-1 levels increase as cells reach senescence. ( A ) Characteristic Immunoblot showing caveolin-1 (Cav-1) levels increasing significantly as WJ-MSCs reach senescence. β-Actin was used as control for equal protein loading. ( B ) Representative photos of early-, middle- and late-passage cells stained with SA-β-gal staining solution. ( C ) Graph showing the percentage of SA-β-gal positive cells in early-, middle- and late-passage cells as mean from three donors. * p < 0.05 vs. NT or otherwise indicated. p values calculated using the Student t -test. ( D ) Representative images of early- and late-passage WJ-MSCs where total ROS and superoxide levels were analyzed under normal conditions (early–late). For a positive control pyocyanine was added to the early passage cells (early + PYO). Images were captured with the 40X objective lens of the fluorescent microscope used.

Article Snippet: Oxidative stress was detected by staining with two fluorescent dyes from the ROS-ID ® Total ROS/Superoxide detection kit (ENZ-51010, Enzo, Farmingdale, NY, USA).

Techniques: Western Blot, Control, Staining, Positive Control, Microscopy

Journal: Cells

Article Title: Dysregulation of Caveolin-1 Phosphorylation and Nuclear Translocation Is Associated with Senescence Onset

doi: 10.3390/cells10112939

Figure Lengend Snippet: Late-passage cells demonstrated an inability to repair DNA damage. ( A ) Characteristic images of early- and late-passage cells where DNA damage was assessed by staining with the 53BP1 antibody and the appropriate secondary antibody (green). (blue). Cav-1 localization was also evaluated by staining with anti-cav-1 antibody and the appropriate secondary antibody (red) under normal conditions (No Treatment—NT) and at 1 h and 24 h after being treated with H 2 O 2 . (300 μΜ, 30 min). Nuclei were stained with DAPI. Images were captured with the 40× objective lens of the fluorescent microscope used. ( B ) Percentages of WJ-MSCs with 53BP1 foci from early- and late-passage cells from three donors evaluated by staining with the 53BP1 antibody under normal conditions (No Treatment—NT) and at different time points of the recovery period from the oxidative treatment (1 h and 24 h). Values shown are the means ± S.E. # p < 0.05 vs. early passage cells at the same time point and * p < 0.05 vs. NT or otherwise indicated. p values calculated using the Student t -test.

Article Snippet: Oxidative stress was detected by staining with two fluorescent dyes from the ROS-ID ® Total ROS/Superoxide detection kit (ENZ-51010, Enzo, Farmingdale, NY, USA).

Techniques: Staining, Microscopy

Journal: Cells

Article Title: Dysregulation of Caveolin-1 Phosphorylation and Nuclear Translocation Is Associated with Senescence Onset

doi: 10.3390/cells10112939

Figure Lengend Snippet: No alterations in Cav-1/pCav-1 levels in late-passage cells after exogenous oxidative insult. ( A ) Representative immunoblot of cell lysates of early- and late-passage WJ-MSCS (from three donors) that have been exposed to H 2 O 2 (300 μΜ, 30 min) and left to recover for 24 h, analyzed at four different time points (NT(=No Treatment), 1 h, 3 h, and 24 h of recovery time post-treatment) by western blotting using the anti-caveolin-1 (Cav-1) antibody. ( B ) Quantification of relative protein expression levels of Cav-1 in cell lysates from early and late WJ-MSCs (from 3 donors) after being exposed to H 2 O 2 was performed based on band density using ImageJ. NT values from both early- and late-passage cells were arbitrarily set to 1. Values shown are the means ± S.E. * p < 0.05 vs. the NT condition. ( C ) Characteristic images of early- and late-passage cells where Cav-1 localization was evaluated by staining with anti-cav-1 antibody and the appropriate secondary antibody (red) under normal conditions (No Treatment—NT) and 24 h after being treated with Doxorubicin. DNA damage was also assessed by staining with the 53BP1 antibody and the appropriate secondary antibody (green). Nuclei were stained with DAPI (blue). Images were captured with the 40× objective lens of the fluorescent microscope used. ( D ) Percentages of WJ-MSCs with 53BP1 foci from early- and late-passage cells from three donors evaluated by staining with the 53BP1 antibody under normal conditions (No Treatment—NT) and 24 h after Doxorubicin treatment. Values shown are the means ± S.E. # p < 0.05 vs. early passage cells at NT and * p < 0.05 vs. NT or otherwise indicated. p values calculated using the Student t -test. ( E ) Representative immunoblot of cell lysates of early- and late-passage WJ-MSCS (from 3 donors) that have been exposed for 24 h to Doxorubicin using anti-caveolin-1 (Cav-1) antibody. ( F ) Quantification of relative protein expression levels of Cav-1 in cell lysates from early and late WJ-MSCs (from 3 donors) after being exposed to 24 h Doxorubicin was performed based on band density using ImageJ. NT values from both early- and late-passage cells were arbitrarily set to 1. Values shown are the means ± S.E. * p < 0.05 vs. the NT condition. ( G ) Representative immunoblot showing p-Cav-1 levels from cell lysates of early- and late-passage WJ-MSCS (from three donors) that have been exposed to H 2 O 2 (300 μΜ, 30 min) and left to recover for 24 h, analyzed at three different time points ((NT(=No Treatment), 1 h and 24 h of recovery time post-treatment). Antibody against β-Actin was used as loading control for all western blots (A, B, D). ( H ) Quantification of relative protein expression levels of p-Cav-1 in cell lysates from early and late WJ-MSCs (from three donors) after being exposed to H 2 O 2 was performed based on band density using ImageJ. Values shown are the means ± S.E. * p < 0.05 vs. the NT condition and # p < 0.05 vs. early passage cells. p values were calculated using Student t -test.

Article Snippet: Oxidative stress was detected by staining with two fluorescent dyes from the ROS-ID ® Total ROS/Superoxide detection kit (ENZ-51010, Enzo, Farmingdale, NY, USA).

Techniques: Western Blot, Expressing, Staining, Microscopy, Control

Journal: Cells

Article Title: Dysregulation of Caveolin-1 Phosphorylation and Nuclear Translocation Is Associated with Senescence Onset

doi: 10.3390/cells10112939

Figure Lengend Snippet: Cav-1 downregulation results in failure of oxidative DNA damage repair. ( A ) Characteristic immunoblot showing caveolin-1 (Cav-1) levels decreasing significantly after siRNA against Cav-1 treatment in early passage cells. Antibody against β-Actin was used as loading control. ( B ) Graph demonstrating Cav-1 levels based on band density calculated using Image J. Values shown are the means ± S.E. * p < 0.05 vs. the early passage WJ-MSCs treated with control siRNA (siCTRL). ( C ) Characteristic images of early passage cells treated with control siRNA and siRNA against Cav-1, under normal conditions (No Treatment—NT) and at 1 and 24 h after being treated with H 2 O 2 (300 μΜ, 30 min) stained with anti-cav-1 antibody and the appropriate secondary antibody (red). DNA damage was also assessed by staining with the 53BP1 antibody and the appropriate secondary antibody (green). Nuclei were stained with DAPI (blue). Images were captured with the 40× objective lens of the fluorescent microscope used. ( D ) Percentages of cells with 53BP1 foci from early passage WJ-MSCs treated with siCTRL and siCav-1 under normal conditions (No Treatment—NT) and at 1 h and 24 h after being treated with H 2 O 2 (300 μΜ, 30 min). Values shown are the means ± S.E. # p < 0.05 vs. siCTRL treated early passage cells and * p < 0.05 vs. NT or otherwise indicated. p values calculated using the Student t -test.

Article Snippet: Oxidative stress was detected by staining with two fluorescent dyes from the ROS-ID ® Total ROS/Superoxide detection kit (ENZ-51010, Enzo, Farmingdale, NY, USA).

Techniques: Western Blot, Control, Staining, Microscopy

Journal: Applications in Plant Sciences

Article Title: Development and characterization of 20 novel EST ‐ SSR markers for Pteroceltis tatarinowii , a relict tree in China

doi: 10.1002/aps3.11320

Figure Lengend Snippet: Characteristics of 20 EST‐SSR markers developed for Pteroceltis tatarinowii .

Article Snippet: For all loci, the 5′ end of each forward primer was tagged with one of four fluorescent dyes (FAM, ROX, HEX, or TAMRA; Sangon Biotech, Shanghai, China).

Techniques: Binding Assay

Journal: Applied spectroscopy reviews

Article Title: Optical spectroscopic imaging for cell therapy and tissue engineering

doi: 10.1080/05704928.2017.1328428

Figure Lengend Snippet: Comparison of physicochemical and optical properties of functional fluorophores: Fluorescent organic dyes, fluorescent proteins, and fluorescent nanoparticles. Modified with permission from Ref. (10). Copyright 2010 SAGE Publications, Inc.

Article Snippet: Copyright 2010 SAGE Publications, Inc. Fluorescent organic dyes The wavelength of fluorophores spans from UV (300–400 nm) and visible spectrum (400– 650 nm) to the NIR spectrum (650–900 nm) ( 12 ).

Techniques: Comparison, Functional Assay, Modification

Journal: Applied spectroscopy reviews

Article Title: Optical spectroscopic imaging for cell therapy and tissue engineering

doi: 10.1080/05704928.2017.1328428

Figure Lengend Snippet: Cell labeling and tracking by molecular imaging modalities. Direct cell labeling utilizes exogenous contrast agents, while indirect labeling is based on genetic modification of cells using fluorescent proteins. Methods for tracking labeled cells include optical imaging, MRI, PET, SPECT, CT, or their combination. Reproduced with permission from Ref. (23). Copyright 2014 Ivyspring International Publisher.

Article Snippet: Copyright 2010 SAGE Publications, Inc. Fluorescent organic dyes The wavelength of fluorophores spans from UV (300–400 nm) and visible spectrum (400– 650 nm) to the NIR spectrum (650–900 nm) ( 12 ).

Techniques: Labeling, Imaging, Modification, Optical Imaging, Single Photon Emission Computed Tomography