Journal: Cells

Article Title: Osteopontin Activation and Microcalcification in Venous Grafts Can Be Modulated by Dexamethasone.

doi: 10.3390/cells12222627

Figure Lengend Snippet: Figure 1. Porcine interposition vein grafts at 7 and 14 days post-surgery were obtained and compared to the control. (A) Immunofluorescence staining with specific antibody for OPN (red) and CD31 (green) as an EC marker was carried out. OPN was quantified in sections and averaged for each experimental group. Representative images, values from four independent experiments, and mean values are shown. (B) Representative images for Von Kossa staining of sections at 7 and 14 days post-surgery compared to the control showing increased stain uptake. (C) Representative images for Alizarin staining of sections at 7 and 14 days post-surgery compared to the control, showing increased stain update. (D) 18F-sodium fluoride uptake at 7 and 14 days post-surgery compared to the control. Representative images, values from four independent experiments, and mean values are shown. p-value * < 0.05, ** < 0.01, *** < 0.001.

Article Snippet: Sections were then incubated in immunofluorescence blocking buffer (Cell Signalling Technology, 12411) for 30 min at room temperature, followed by overnight incubation in primary antibody (Table 1) and immunofluorescence antibody dilution buffer (Cell Signalling Technology, 12378) at 4 ◦C.

Techniques: Control, Staining, Marker

Journal: Cells

Article Title: Osteopontin Activation and Microcalcification in Venous Grafts Can Be Modulated by Dexamethasone.

doi: 10.3390/cells12222627

Figure Lengend Snippet: Figure 3. LSV were either pretreated with dexamethasone for 60 min or remained untreated. They were then mounted on a perfusion apparatus and exposed to LSS for 4 h or 45 min. (A) Transcript levels of MCP-1 and IL-8 were measured by comparative RT-PCR. Values from four independent experiments with veins either pretreated with dexamethasone or kept untreated as control and mean values are shown. (B) Immunofluorescence staining with specific antibody for active (phospho) P38 (red) and CD31 (green) as an EC marker was carried out with and without dexamethasone pretreatment. (C) Transcript levels of OPN were measured by comparative RT-PCR. Values from four independent experiments with veins either pretreated with dexamethasone or kept untreated as a control and mean values are shown. (D) RNAscope with a probe specific for the OPN gene (arrows indication expression of genes) quantified in multiple sections and averaged for each experimental

Article Snippet: Sections were then incubated in immunofluorescence blocking buffer (Cell Signalling Technology, 12411) for 30 min at room temperature, followed by overnight incubation in primary antibody (Table 1) and immunofluorescence antibody dilution buffer (Cell Signalling Technology, 12378) at 4 ◦C.

Techniques: Reverse Transcription Polymerase Chain Reaction, Control, Staining, Marker, RNAscope, Expressing

Journal: Cells

Article Title: Osteopontin Activation and Microcalcification in Venous Grafts Can Be Modulated by Dexamethasone.

doi: 10.3390/cells12222627

Figure Lengend Snippet: Figure 4. LSV were either pretreated with dexamethasone (10 µM for 60 min) or remained untreated. They were then mounted on a perfusion apparatus and exposed to LSS for 4 h then cultured in a culture well for 7 and 14 days. (A) Immunofluorescence staining with specific antibody for OPN (red) and CD31 (green) as an EC marker was carried. OPN was quantified in sections and averaged for each

Article Snippet: Sections were then incubated in immunofluorescence blocking buffer (Cell Signalling Technology, 12411) for 30 min at room temperature, followed by overnight incubation in primary antibody (Table 1) and immunofluorescence antibody dilution buffer (Cell Signalling Technology, 12378) at 4 ◦C.

Techniques: Cell Culture, Staining, Marker

Journal: Journal of Thoracic Disease

Article Title: ZLN005 improves the protective effect of mitochondrial function on alveolar epithelial cell aging by upregulating PGC-1α

doi: 10.21037/jtd-23-815

Figure Lengend Snippet: Determination of PGC-1α, NRF-1 protein, mitochondrial morphology, and function, and the expression levels of cell senescence-related markers (i.e., p21 WAF1 and SA-β-gal) in the H 2 O 2 -induced AEC senescence model. The A549 cells and mice AEC2 cells were cultured in serum-free medium and treated with different concentrations of H 2 O 2 for the following experiments: (A) Western blot was used to detect the expression levels of PGC-1α, NRF-1, and p21 WAF1 in the AECs; (B) a quantitative analysis of the Western blot experiment results was conducted; (C,D) SA-β-gal staining of the A549 and AEC2 cells treated with different concentrations of hydrogen peroxide was performed (×100); (E,F) the positive rates of the SA-β-gal staining in the A549 and AEC2 cells was determined; (G) immunofluorescence experiments were conducted to determine the mitochondrial morphological changes in the A549 cell (Mitotracker Red, ×800); and (H,I) immunofluorescence experiments were conducted to determine of the mitochondrial morphological changes and the expression levels of p21 WAF1 in the AECs (×200). β-actin served as an internal parameter. *, P<0.05; **, P<0.01; ***, P<0.001; ns, no significance. PGC-1α, peroxisome proliferator-activated receptor-γ coactivator-1 alpha; NRF-1, nuclear respiratory factor-1; DAPI, 4',6-diamidino-2-phenylindole; SA-β-gal, senescence-associated beta-galactosidase; AEC, alveolar epithelial cell.

Article Snippet: The immunofluorescence kit (Cell Signaling Technology, Boston, USA) was used for the immunofluorescence analysis of the antibodies against p21 in accordance with the manufacturer’s instructions.

Techniques: Expressing, Cell Culture, Western Blot, Staining, Immunofluorescence