Journal: Journal of Biological Chemistry

Article Title: Differential Regulation of NOTCH2 and NOTCH3 Contribute to Their Unique Functions in Vascular Smooth Muscle Cells

doi: 10.1074/jbc.m115.655548

Figure Lengend Snippet: FIGURE 5. NOTCH3 protects smooth muscle cells from cell death. HAoSMCs with lentivirally overexpressed NICD2, NICD3, and GFP, or siRNA knockdown of NOTCH2 (siN2), NOTCH3 (siN3), and control (siCTL) were exposed to 50 J/cm2 UV radiation and collected for protein 9 h later. A and B, Western blots to detect NOTCH2 and NOTCH3 expression, and level of total and cleaved caspase3 with tubulin used as loading control, n 3. C and D, caspase3 enzymatic activity assay using fluorescent caspase substrate AFC-DEVD, n 4. Significance determined by one-way ANOVA; *, p 0.05.

Article Snippet: Lysates were then used in a caspase3 enzymatic activity assay using AFCDEVD substrate (MP Biomedicals) as described previously (43).

Techniques: Knockdown, Control, Western Blot, Expressing, Enzyme Activity Assay

Journal: Journal of Biological Chemistry

Article Title: Differential Regulation of NOTCH2 and NOTCH3 Contribute to Their Unique Functions in Vascular Smooth Muscle Cells

doi: 10.1074/jbc.m115.655548

Figure Lengend Snippet: FIGURE 7. Genetic deletion of Notch3 has unique effects on the survival of mouse aortic smooth muscle cells. A, mRNA expression of pro-survival genes in aortas of wild-type (WT), Notch2-deficient (Notch2fl/fl; MCC/), and Notch3-mutant (Notch3/) adult mice. qPCR with relative expression compared with GAPDH, n 4. B, SMCs were isolated and cultured from aortas of wild-type, Notch2, and Notch3-deficient mice and used in enzymatic caspase3 activity assays, n 4. C, total and phosphorylated (phospho)-ERK was measured in cultured smooth muscle cells following serum challenge, with tubulin used as loading control, n 4. Significance determined by one-way ANOVA, *, p 0.05.

Article Snippet: Lysates were then used in a caspase3 enzymatic activity assay using AFCDEVD substrate (MP Biomedicals) as described previously (43).

Techniques: Expressing, Mutagenesis, Isolation, Cell Culture, Activity Assay, Control

Journal: PLoS Genetics

Article Title: Cdh11 Acts as a Tumor Suppressor in a Murine Retinoblastoma Model by Facilitating Tumor Cell Death

doi: 10.1371/journal.pgen.1000923

Figure Lengend Snippet: Antibody list.

Article Snippet: Activated Caspase-8 , Abnova , 1∶1000.

Techniques: Purification

Journal: Scientific Reports

Article Title: The VP3 protein of duck hepatitis A virus mediates host cell adsorption and apoptosis

doi: 10.1038/s41598-019-53285-0

Figure Lengend Snippet: Detection results of apoptotic caspase-3, −8 and −9 transcriptional levels and enzyme activities. ( A ) Detection of transcript levels of apoptotic caspase-3, −8 and −9 in pCAGGS/VP3-transfected DEFs. ( B ) Detection of enzymatic activity of apoptotic caspase-3, −8 and −9 in pCAGGS/VP3-transfected DEFs.

Article Snippet: DEFs were transfected with pCAGGS/VP3 or pCAGGS for 48 h. In a 96-well plate, 100 μL of a cell (1~5 × 104 cells) suspension and 100 μL of caspase-3, caspase-8 and caspase-9 enzyme activity detection reagent (Promega) was added separately and mixed; the cells were incubated at 22 °C for 1 h in the dark and analyzed using a multifunction microplate reader within 3 h.

Techniques: Transfection, Activity Assay