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. 2021 Apr 9;36(5):890–900. doi: 10.1007/s12250-021-00369-9

Fig. 6.

Fig. 6

Construction a SARS-CoV-2-GFP-FLuc replicon and determination of the EC50 of E64-D. A Diagram of SARS-CoV-2-GFP-FLuc replicon. B Verification of a SARS-CoV-2-GFP-FLuc replicon plasmid. Recombinant SARS-CoV-2-GFP-FLuc replicon plasmid was tested for all sub-genomic fragments used in yeast homologous recombination by PCR. C Luciferase activity in nontransfected (BLANK) or SARS-CoV-2-GFP-FLuc replicon-transfected or 293 T cells, the luciferase activity was determined 36 h post transfection. D Kinetics study of luciferase activity after SARS-CoV-2-GFP-FLuc replicon transfection, 293 T cells were transfected with SARS-CoV-2-GFP-Fluc replicon and luciferase activity was determined at indicated hours post-transfection (hpt). E EC50 value of E-64D was determined using a dose response curve drawn based on luciferase activity in SARS-CoV-2-GFP-FLuc-transfected 293 T cells using four-parameter nonlinear regression. F EC50 value of E-64D was determined using a dose response curve drawn based on the ratio of tGFP-BlaR/β-actin gray values with different concentration and calculated by four-parameter nonlinear regression, the experiments were done at least two times.