RT-PCR and 2-D western blot analysis of EGFP, 3CLpro, and C145A transfected A549 cell line. (A) RT-PCR analysis of 3CLpro and GM-CSF genes expression at 12, 24, and 48 hrs in A549 cells which were transfected with EGFP, 3CLpro, and C145A respectively. The relative abundance compared with the expression level of GAPDH is depicted in the bar chart below. (B) 2-D western blots of 3CLpro and C145A expression patterns in A549 cells. A549 cells transfected with EGFP, 3CLpro and C145A were analyzed at 24 and 48 hrs after transfection. Proteins were resolved by 2-D gels and electroblotted onto PVDF membranes. Western blots were probed with anti-3CLpro specific antibody. The circle indicates the site of 3CLpro.