Journal: Acta biochimica et biophysica Sinica

Article Title: Monocyte chemoattractant protein-1 induces endothelial cell apoptosis in vitro through a p53-dependent mitochondrial pathway.

doi: 10.1093/abbs/gmr072

Figure Lengend Snippet: Figure 1 MCP-1 induced HUVECs apoptosis (A) HUVEC strain CRL-1730 (ATCC) was cultured in complete DMEM/F12 culture medium (containing 20% FBS) for 24 h. The cells appeared uniform ECs characteristic ‘cobble stone’ morphology. Original magnification 100. (B) The cultured cells were immunofluorescent positive for EC-specific marker vWF. Original magnification 300. (C) MTT assay indicated that MCP-1 inhibited HUVECs proliferation dose dependently. (D) After treatment with MCP-1(0.1–100 ng/ml) for 48 h, apoptosis rate was detected. MCP-1 increased HUVECs early apoptotic (FITCþ/PI2) rates as compared with Cont. Cont, cells not treated with MCP-1. *P , 0.05, **P , 0.01 compared with the Cont. Data were expressed as mean+SD from three separate experiments.

Article Snippet: NAO (10-N-nonyl acridine orange) is a derivative of acridine Figure 1 MCP-1 induced HUVECs apoptosis (A) HUVEC strain CRL-1730 (ATCC) was cultured in complete DMEM/F12 culture medium (containing 20% FBS) for 24 h. The cells appeared uniform ECs characteristic ‘cobble stone’ morphology.

Techniques: Cell Culture, Marker, MTT Assay

Journal: Acta biochimica et biophysica Sinica

Article Title: Monocyte chemoattractant protein-1 induces endothelial cell apoptosis in vitro through a p53-dependent mitochondrial pathway.

doi: 10.1093/abbs/gmr072

Figure Lengend Snippet: Figure 3 Mitochondria involvement in MCP-1-induced HUVECs apoptosis After HUVECs were treated with MCP-1 (0.1–100 ng/ml) for 48 h, indicators for mitochondrial membrane potential (Dcm) and mitochondrial mass, and labeled cyt c antibody were separately loaded for following flow cytometry. (A) Rh123 fluorescence reduction indicated that HUVECs mitochondrial membrane potential was decreased by MCP-1 treatment. (B) NAO fluorescence reduction implied that MCP-1 treatment resulted in decrease of HUVECs mitochondrial mass. (C) Mitochondrial cyt c fluorescence decreased and meanwhile cytoplasmic cyt c fluorescence increased, indicating that MCP-1 promoted HUVECs cyt c release. Cont, cells not treated with MCP-1. *P , 0.05, **P , 0.01 compared with the Cont. Data were expressed as mean + SD from three separate experiments.

Article Snippet: NAO (10-N-nonyl acridine orange) is a derivative of acridine Figure 1 MCP-1 induced HUVECs apoptosis (A) HUVEC strain CRL-1730 (ATCC) was cultured in complete DMEM/F12 culture medium (containing 20% FBS) for 24 h. The cells appeared uniform ECs characteristic ‘cobble stone’ morphology.

Techniques: Membrane, Labeling, Flow Cytometry, Fluorescence

Journal: Acta biochimica et biophysica Sinica

Article Title: Monocyte chemoattractant protein-1 induces endothelial cell apoptosis in vitro through a p53-dependent mitochondrial pathway.

doi: 10.1093/abbs/gmr072

Figure Lengend Snippet: Figure 4 p53 played an important role in MCP-1-induced HUVECs apoptosis After treated with MCP-1 (0.1–100 ng/ml) for 48 h, p53 mRNA (A) and protein (B) levels were both found to be upregulated. Cont, cells not treated with MCP-1. *P , 0.05, **P , 0.01 compared with the Cont. (C) HUVECs were pretreated with pifithrin-a (10–30 mM) for 30 min and followed by 10 ng/ml MCP-1 treatment for 48 h. HUVECs apoptosis was only rescued by 30 mM pifithrin-a as revealed by Annexin V-FITC/PI double-staining flow cytometry. *P , 0.05 compared with cells treated with 10 ng/ml MCP-1 alone. Data were expressed as mean+SD from three separate experiments.

Article Snippet: NAO (10-N-nonyl acridine orange) is a derivative of acridine Figure 1 MCP-1 induced HUVECs apoptosis (A) HUVEC strain CRL-1730 (ATCC) was cultured in complete DMEM/F12 culture medium (containing 20% FBS) for 24 h. The cells appeared uniform ECs characteristic ‘cobble stone’ morphology.

Techniques: Double Staining, Flow Cytometry

Journal: Acta biochimica et biophysica Sinica

Article Title: Monocyte chemoattractant protein-1 induces endothelial cell apoptosis in vitro through a p53-dependent mitochondrial pathway.

doi: 10.1093/abbs/gmr072

Figure Lengend Snippet: Figure 5 PKC modulated MCP-1-induced HUVECs apoptosis (A) MCP-1(10 ng/ml) stimulation for 48 h augmented intracellular calcium ([Ca2þ]i) concentrations. Cont, cells not treated with MCP-1. *P , 0.05, **P , 0.01 compared with the Cont. After being treated with different concentrations of PKC activator PMA (0.01–1 mM) (B) or PKC inhibitor chelerythrine (0.1–10 nM) (C) for 12 h, HUVECs were induced by 10 ng/ml MCP-1 for 24 h, and then the apoptosis rates were detected by using FCM. Cont, cells not treated with MCP-1. *P , 0.05, **P , 0.01 compared with the cells treated with 10 ng/ml MCP-1 alone. Che, chelerythrine. Data were expressed as mean+SD from three separate experiments.

Article Snippet: NAO (10-N-nonyl acridine orange) is a derivative of acridine Figure 1 MCP-1 induced HUVECs apoptosis (A) HUVEC strain CRL-1730 (ATCC) was cultured in complete DMEM/F12 culture medium (containing 20% FBS) for 24 h. The cells appeared uniform ECs characteristic ‘cobble stone’ morphology.

Techniques: