FIG 5.

Inhibition of CPT1a increases MDV titer. (A) Fold-change expression of genes involved in fatty acid oxidation in MDV-infected CEFs at 24, 48, and 72 hpi. (B) Hydrogen peroxide (H₂O₂) synthesis (μM) in mock- or MDV-infected CEFs treated with etomoxir (4.42 μM) or clofibrate (0.41 μM) at 72 hpi. Analysis of MDV viral titer in MDV-infected CEFs treated with (C) clofibrate (0.01, 0.02, 0.041, 0.1, 0.2, and 0.41 μM), an agonist of PPAR-α, (D) malonyl-CoA (5, 10, 15, 25, and 30 μM), and (E) etomoxir (0.29, 1.47, 2.95, and 4.42 μM). (F) MDV viral titers are shown for MDV-infected CEFs treated with etomoxir (4.42 μM) or in combination with either palmitic acid (5, 12.5, and 25 μM) or malonyl-CoA (5, 15, and 25 μM). (G) MDV genome copy numbers per 10⁴ cells (meq gene with reference ovotransferrin gene) were determined using qPCR in CEFs treated with etomoxir (4.42 μM) or clofibrate (0.41 μM). Nonparametric Wilcoxon tests (Mann-Whitney) and one-way ANOVA were used to assess normal distribution and test significance with the results shown as mean ± SD. * (P < 0.05) and **** (P < 0.0001) indicate a statistically significant difference compared to the control. NS indicates no significant difference. All experiments were performed in 6 replicates for plaque assays and 3 replicates for real-time PCR and fluorometric assays. All experiments were performed in triplicate, and data are representative of 3 independent experiments.