Fig 6. Inhibition of PC synthesis restores neutral lipid synthesis in infected cells but does not significantly affect the first cycle of poliovirus replication.

A. Scheme of the experiments shown on panels B-D. HeLa cells pre-incubated in choline-free medium for ~72h were infected with poliovirus and were incubated after infection either in choline-free or choline-supplemented medium. B. Incorporation of the fluorescent long chain FA analog added to choline-deprived HeLa cells infected with an MOI of 10 PFU/ml of poliovirus at 4 h p.i. for 1 h. In cells incubated in choline-free medium, it redistributes to neutral lipids in lipid droplets (left panel), while in cells incubated with choline-supplemented medium it is used for membrane synthesis (right panel). C. Accumulation of the viral proteins 2C and 2BC (left panel) at 4 h p.i. and total virus yield (right panel) at 6 h p.i. did not depend on whether previously choline-deprived cells were incubated in choline-free or choline-supplemented medium after infection. Actin shown as a loading control. D. Total virus yield at 6 h p.i. upon infection of choline-deprived cells at low MOIs of 0.05 and 0.5 does not depend on whether the cells were incubated in choline-free or choline-supplemented medium after infection. E. Presence of choline in the incubation medium also did not affect the dynamics of accumulation of extracellular (extra) and intracellular (intra) progeny virus upon infection of choline-deprived HeLa cells. Virus yield at the indicated time points was determined after infection of choline-deprived HeLa cells with an MOI of 10 and incubation after infection in choline-free or choline-supplemented media.