Ecl plus a and b

Protein synthesis rates were measured using a nonradioactive method [67 (link)]. Puromycin (10 μM; Solarbio, Beijing, CN) was added to cell culture media for 30 min after the addition of the treatments listed above, and total proteins were extracted and used to measure protein synthesis rates. Newly synthesized polypeptides were labelled with Puromycin at low concentrations to reflect the rate of protein synthesis [67 (link), 68 (link)]. The protein-antibody complexes were detected with ECL Plus A and B (Beyotime, Nanjing, Jiangsu, CN), and the results were quantified using the Fusion FX software (Vilber Lourmat, Paris, FR).

The protein synthesis rate was detected using a nonradioactive method [28 (link)]. The newly synthesized proteins labeled with puromycin were subsequently detected with an anti-puromycin antibody. The accumulation of puromycin-conjugated peptides into nascent peptide chains reflects the rate of protein synthesis [28 (link), 29 (link)]. The protein-antibody complexes were detected with ECL Plus A and B (Beyotime, Nanjing, Jiangsu, CN), and the results were quantified using the Fusion FX software (Vilber Lourmat, Paris, FR).