HepG2 cell APOBEC3B-AcGFP anti-AcGFP-IP DIA analysis

APOBEC3B-AcGFP expressing HepG2 or AcGFP expressing HepG2 plated on 10 cm dishes were stimulated with Act D (1 µM), MNNG (50 µM) for 3 h. The cells were washed with PBS three times and lysed with RIPA buffer containing protease inhibitor cocktail. The protein amounts of cell lysates were measure in BCA protein assay. 0.1 mL of the cell lysate (5 mg/mL) were added with 4 µL of Anti GFP (VHH), GFP-Trap Magnetic Agarose and rotated for 1 h at 4˚C. The obtained data were analyzed using DIA-NN 1.8.1. Proteins with both precursor FDR and protein FDR of 1% were identified and quantification values were calculated.