FIG 5.

AMPK inhibitor and agonists affect viral lytic replication but have no effect on virus entry, trafficking, and infectivity during KSHV primary infection. (A) Effects of AMPK inhibitor and agonists on AMPK activity. HUVEC were treated with compound C (Com C, 5 μM), AICAR (1 mM), or metformin (Met, 1 mM) for 8 h and examined for phosphorylated AMPK (T172) (p-AMPK), total AMPK, phosphorylated ACC1 (S79) (p-ACC1), and total ACC1. NT, no treatment. (B and C) AMPK inhibitor increases, while agonists inhibit the production of infectious virions. HUVEC pretreated with AMPK inhibitor or agonists for 1 h were infected with KSHV in the presence of the chemicals for 24 h. Cells were extensively washed, and medium was replaced. At day 4 postinfection, the supernatants were collected and subjected to titration of infectious virions as described for Fig. 2E and F. (D and E) AMPK inhibitor and agonists have no effect on virus entry and trafficking. HUVEC pretreated with AMPK inhibitor or agonists for 1 h were infected with KSHV for 6 h in the presence of the chemicals. The number of KSHV particles successfully docked at the nuclei was analyzed as described for Fig. 2G and H. (F and G) AMPK inhibitor and agonists have no effect on the expression level and pattern of KSHV latent protein LANA or infectivity. HUVEC pretreated with AMPK inhibitor or agonists for 1 h were infected with KSHV in the presence of these chemicals for 48 h and analyzed for the expression level and pattern of LANA (F) and the numbers of LANA-positive cells (G).