MAP-Kinase Regulated Cytosolic Phospholipase A2 Activity Is Essential for Production of Infectious Hepatitis C Virus Particles
Figure 4
Knock down of PLA2G4A increases susceptibility of HCV to Py-2.
Huh-7.5 cells were transfected with PLA2G4A-specific or scrambled siRNAs. 96 h later cells were inoculated with a concentrated Luc-Jc1 virus preparation (A) Efficiency of PLA2G4A knock down was controlled by Western blot. (B) PLA2G4A enzyme activity was measured using a commercial assay. (C) HCV RNA replication was quantified in cell lysates (top panel) and release of infectious particles was determined after inoculation of naïve Huh-7.5 cells (bottom panel). Data are shown as means +/− SD of three independent experiments (the dotted line represents background luciferase activity in mock infected cells). (D and E) Huh-7.5 cells were transfected with PLA2G4A-specific or scrambled siRNAs. 96 h later, cells were infected with Luc-Jc1 particles and subjected to the inhibitor assay outlined in Figure 1A. (D) Knock down of PLA2G4A was monitored by Western blotting. (E) HCV RNA replication was measured in cell lysates (top panel) and release of infectious particles was determined by inoculation of naïve Huh-7.5 cells (bottom panel) and luciferase assays. The box plots in panels (B) and (E) as well as in the following figures visualize the full distribution of the data; the central horizontal line in each box indicates the value of the median, whereas the box represents the range between the lower and upper quartile of the data, i.e. the area in which the central 50% of the measurements lie. The whiskers extend from the quartiles to the minimum and maximum measurements, respectively. Statistical significance of difference between means is indicated using asterisks (*): n.s - not significant, * marginally significant (p≤0.1), ** significant (p≤0.05), *** highly significant (p≤0.01).