Fig. 7.

2-DG treatment produces defective progeny virions.

A. The graphical depiction indicates the methodology followed for the experiment of progeny virion (P3) infection. The cells were infected with the normalized concentration of progeny virions, and no further 2-DG treatment was given following infection. B. The photomicrographs show the progeny virions induced CPE by bright field imaging in the indicated treatment groups at 48 h post-infection. C. The estimation of N and RdRP genes was performed in the P3 infected samples by RT-PCR assay (n = 3; from three independent observations). Initially, the P3 virions in the supernatant were quantified by RT-PCR from infected, and 2-DG treated cells, and equal virion titer was obtained by volume-dependent concentration normalization depending upon the respective Ct values. Finally, Vero E6 cells were infected with these normalized concentrations of virions from infected and 2-DG treated (1 & 5 mM) groups. D. Immunoblotting of virus N and S protein was performed in cell lysate infected with P3 progeny virion of indicated treatment groups at 12 h post-infection (additional blot images are also provided in supplementary Fig. 5B); The densitometry graph shows the RFC in N and S proteins (from three and two independent observations respectively) among indicated progeny fractions. E. Crystal-violet stained plaques are presented as the plaque-forming unit (PFU)/ml of progeny titer in the indicated treatment groups where Vero E6 cells were infected with the equal concentration of P3 progeny virions from the infected and 2-DG treated (0.5, 1 & 5 mM) groups using 10-fold serial dilution (n = 3). F. The barplot indicates the quantitative estimation of plaques (derived from Fig. 7 E; from two independent observations) in the indicated treatment groups. Data represent the mean ± SD and P-values in all the barplots calculated with respect to the infected group using the two-tailed Student's t-test. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)