Figure 7.

Pretreatment with Compound 22 reduces RVFV replication. (a) HSAECs were pretreated with 2 and 4 µM of Compound 22 or DMSO only in conditioned media for one hour. Following pretreatment, cells were infected with rMP12 at MOI 5 for one hour. Cells were washed once in PBS, then treated, conditioned media containing the inhibitor was replaced for the remaining time. At 24 hpi, cell culture media was removed and lysates were collected and analyzed by western blot. Membranes were probed for p-Akt (Ser 473), pan-Akt and β-Actin as a loading control. Cropped, representative images are shown here. Uncropped membranes can be found in Supplemental Fig. 8. (b) Vero cells and HSAECs were pretreated with Compound 22 (5 µM or 1 µM) or DMSO for one hour. Following treatment, cells were mock-infected or infected with MP12-Luciferase at MOI 0.1 for one hour. Cells were washed, then complete media was added. At 24 hpi, replication was measured by Promega’s Renilla Glo Luciferase assay according to manufacturer’s protocol. (c) Vero cells or HSAECs were pretreated and infected as in Panel B, then supernatants were collected at 24 hours post infection and analyzed by plaque assay. Data are expressed as the Mean ± SD (n = 3). *p ≤ 0.05; ****p ≤ 0.0001