Figure 1

Reverse PCR transcription analysis and dynamics of eGFP-positive cell accumulation in cultures. A. Transcripts were detected 48 h after A549 and Calu-1 transfection with pBI-EGFP (Mock), pBI-EGFP/3C, encoding intact 3C protease (3Cpro), and pBI-EGFP/3CMut, encoding catalytically inactive 3C protease (3CMut). Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a reference gene. B. The proportions of eGFP-positive cells were counted in A549 and Calu-1 cultures 144 h post-transfection (p.t.) with pBI-EGFP/3C (black points, solid line) and pBI-EGFP (light points, dotted line). Presented values were averaged for at least two independent experiments including four measurements each. The confidence interval was calculated at 95%. C. 3Cpro-expressing cells 48 h p.t. Normal (N), cells with normal morphology; Rounded/Blebbed (R/B), round and shrunk cells with smooth or blebbed plasma membrane; Vacuolated (V), vacuolated cells. D. Proportions of eGFP-positive cells of different morphology in control (gray bars) and experimental (empty bars) cultures. The values were averaged for at least two independent experiments. Two hundred cells were counted in three replicates for each time point. The confidence interval was calculated at 95%.