Journal: Scientific Reports

Article Title: MicroRNA410 Inhibits Pulmonary Vascular Remodeling via Regulation of Nicotinamide Phosphoribosyltransferase

doi: 10.1038/s41598-019-46352-z

Figure Lengend Snippet: miR410 overexpression increases hPAECs apoptosis. ( A ) Representative images of TUNEL positive cells (red), nucleus was stained by DAPI, and percentage of TUNEL positive cells was quantified in panel B. *P < 0.05, ***P < 0.001 relative to control; ### P < 0.001 relative to 50 nM group. Size bar: 20 µm.

Article Snippet: Three groups of cells cultured in 6-well plates were treated as described in the mitochondrial depolarization assay, then fixed with 4% paraform phosphate buffer saline, rinsed with PBS, then used in situ BrdU-Red DNA fragmentation (TUNEL) cell apoptosis detection kit (Abcam®, Cambridge, MA).

Techniques: Over Expression, TUNEL Assay, Staining

Journal: FASEB BioAdvances

Article Title: Cyclophilin A induces macrophage apoptosis and enhances atherosclerotic lesions in high‐fat diet‐fed hyperglycemic rabbits

doi: 10.1096/fba.2020-00135

Figure Lengend Snippet: Cyclophilin A increases macrophage inflammation and apoptosis in the lesion area of HFD‐fed rabbits through caspase 3 mediated cell apoptosis. (A i) TUNEL staining showing increased apoptotic cell population in the aortic lumen of HFD‐fed rabbit compared to the aortic lumen of the ND fed rabbit; Scale bar: 10 µm (ii) Quantitative data revealing that HFD‐fed rabbit aortas have a significant increase in apoptotic cells in the lesion area. The data are expressed as the percentage of TUNEL‐positive cells per field (four fields per two rabbits from each group). (B) Immunoblotting assay: Protein level expression of Cytochrome C and Cleavage of caspase 3 were increased in aortic tissue of HFD‐fed rabbit compared to ND fed rabbit. (C i) Immunostaining images for M3/84 (green) and Dapi (blue) in aorta from New Zealand White rabbits fed with HFD for 12 weeks (n = 12) compared with ND fed rabbit ( n = 12) demonstrating macrophage inflammation in aorta intima. Scale bar: 10 µm. (C ii) Quantitative data revealing that HFD‐fed rabbit aortas have a significant increase in inflammatory macrophages in the lesion area. * p < 0.05 vs. ND, ** p < 0.01 vs. ND, *** p < 0.001 vs. ND. The data were compared using a one‐way ANOVA followed by multiple comparison tests using the SNK test. Results are the mean ±SD and show pooled data from two experiments (ND, Normal Diet; HFD, High‐Fat Diet; TUNEL, terminal deoxynucleotidyl transferase dUTP nick end labeling, M3/84 ‐ LAMP‐2 Antibody; SNK test, Student–Newman–Keuls test)

Article Snippet: Apoptosis was quantified using TUNEL Assay Kit ‐ In situ BrdU‐Red DNA Fragmentation (Abcam; ab‐66110) according to manufactures description.

Techniques: TUNEL Assay, Staining, Western Blot, Expressing, Immunostaining, Comparison

Journal: FASEB BioAdvances

Article Title: Cyclophilin A induces macrophage apoptosis and enhances atherosclerotic lesions in high‐fat diet‐fed hyperglycemic rabbits

doi: 10.1096/fba.2020-00135

Figure Lengend Snippet: Cyclophilin A induces apoptosis of high glucose primed macrophages. THP 1 monocytes were cultured with PMA to differentiate into macrophages and treated with cyclophilin A (150 ng/ml) for 24H or siRNA or staurosporine (2 µM for 3H). (A) Cyclophilin A‐induced apoptosis was detected with annexin V FITC PI staining using flow cytometry. FACS dot plot shows necrotic (Q1), early apoptotic cells (Q2), viable (Q3) and late apoptotic (Q4) cell population. (B) Confocal images of early (green) and late (red and green) apoptotic cells treated with cyclophilin A (150 ng/ml) using AnnV/PI staining. Scale bar: 10 µm. (C) DNA fragmentation assay using agarose gel electrophoresis showing cleavage of chromosomal DNA after cyclophilin A treatment for 24H in NG and HG treated cells; # p < 0.05 vs. ND; * p < 0.05 vs. HG, ** p < 0.01 vs. HG, *** p < 0.001 vs. HG The data were compared using a one way ANOVA followed by multiple comparison test using SNK test (PMA, Phorbol 12‐Myristate 13‐Acetate; 24H, 24 hours; siRNA, Small interfering RNA; FITC, Fluorescein isothiocyanate; PI, Propidium iodide; FACS, Fluorescence‐activated cell sorting; NG, Normal Glucose; HG, High Glucose; SNK test, Student–Newman–Keuls test)

Article Snippet: Apoptosis was quantified using TUNEL Assay Kit ‐ In situ BrdU‐Red DNA Fragmentation (Abcam; ab‐66110) according to manufactures description.

Techniques: Cell Culture, Staining, Flow Cytometry, DNA Fragmentation Assay, Agarose Gel Electrophoresis, Comparison, Small Interfering RNA, Fluorescence, FACS

Journal: FASEB BioAdvances

Article Title: Cyclophilin A induces macrophage apoptosis and enhances atherosclerotic lesions in high‐fat diet‐fed hyperglycemic rabbits

doi: 10.1096/fba.2020-00135

Figure Lengend Snippet: Cyclophilin A induces apoptosis of high glucose activated macrophages through mitochondria‐mediated intrinsic death signaling pathways. (A) High glucose induces ROS in the aortic vascular lumen which increases secretion of cyclophilin A by activated monocytes leading to increased uptake of OxLDL by overexpressing CD 36. (B) Intracellular cyclophilin A increases mitochondrial ROS resulting in both mitochondrial membrane potential loss and mPTP opening leading to the induction of cytochrome C and activation of caspase 3. (C) Intracellular cyclophilin A induces chromosomal DNA fragmentation in the presence of high glucose. (D) Both the nuclease activity of cyclophilin A and the induction of mitochondria‐mediated death signaling pathways induce macrophage apoptosis (ROS‐ Reactive oxygen species; OxLDL‐ Oxidized low‐density lipoprotein; CD36‐ cluster of differentiation 36; mPTP‐ mitochondrial permeability transition pore).

Article Snippet: Apoptosis was quantified using TUNEL Assay Kit ‐ In situ BrdU‐Red DNA Fragmentation (Abcam; ab‐66110) according to manufactures description.

Techniques: Protein-Protein interactions, Membrane, Activation Assay, Activity Assay, Permeability