Bca protein assay reagent

Caspase colorimetric assay kits (Keygen Biotech, China) were used to measure the activity levels of caspase-3, caspase-8 and caspase-9. EECs were pre-treated with rapamycin, NH₄Cl, chloroquine and wortmannin for 6 h and transfected with shRNA prior to viral infection; then, the cells were infected with PPRV at an MOI of 1. At the indicated time points, the cells were treated with lysis buffer, and the protein concentrations were measured using BCA protein assay reagent (Vazyme, NJ, USA). Then, 150-μg lysates of each sample were loaded into microplates and incubated with each caspase substrate at 37°C for 4 h; after that, the absorbance values of the samples were measured at 405 nm in a microplate spectrophotometer (Infinite 200 PRO NanoQuant, Tecan, Switzerland).

The activities of caspase-3, -8, and -9 were detected by colorimetric assay kit (KeyGEN Biotech). The cells were incubated with lysis buffer, and the concentrations of protein were detected by bicinchoninic acid (BCA) protein assay reagent (Vazyme Biotech, Nanjing, Jiangsu Province, China). The protein (200 μg/sample) was treated with caspase-3, -8, and -9 substrate for each sample at 37 °C for 4 h. Samples were read by a microplate reader (model 680, Bio-Rad) at 405 nm.

Caspases activities were measured by colorimetric assay kits (Keygen Biotech, Nanjing, China) following the manufacture’s protocol. Briefly, protein concentrations were measured by BCA Protein Assay Reagent (Vazyme Biotech Nanjing, China). Then 200 mg protein of each sample was incubated with each caspase substrate at 37°C in a microplate for 4 h. Absorbance at the wavelength of 405 nm was read in microplate spectrophotometer (Infinite 200 PRO NanoQuant; Tecan, Switzerland).