Figure 7. Autophagy Inducers Inhibit RVFV Infection in Primary Mouse Hepatocytes and Rat Neurons.

(A) Primary mouse hepatocytes were infected with RVFV (MOI 5) and protein was isolated at the indicated time points after infection to assess p62 and LC3 expression by immunoblot. Data are representative of two independent experiments.

(B) RVFV RNA levels from primary mouse hepatocytes pretreated with SMER28 or rapamycin and infected with RVFV (MOI 1) for 20 hr as determined by qRTPCR. Data are representative of two independent experiments.

(C) Isg56 mRNA expression quantified by qRT-PCR from RVFV-infected mouse hepatocytes treated with the indicated drugs or vehicle control at 20 hpi.

(D) Primary rat neuronal cultures were infected with RVFV for the indicated times (MOI 1) or treated with rapamycin (1 μM) or SMER28 (50 μM) for 16 hr. Immunoblot analysis of LC3 and p62 expression is shown. Data are representative of two independent experiments.

(E) Primary rat neuronal cultures were pretreated with SMER28 or vehicle control for 1 hr and infected with RVFV (MOI 0.1) for 22 hr. Infection was assessed by immunofluorescence.

(F) Fold decrease in the percent of RVFV-infected rat neuronal cells with SMER28 treatment. Mean ± SE for three independent experiments; *p < 0.05, Student's t test.

(G) Immunofluorescence of RVFV and neurons (MAP2-positive cells) from RVFV-infected neuronal cultures pretreated with SMER28 or vehicle control showing preferential protection of neurons with SMER28 treatment.

(H) RVFV RNA was quantified by qRT-PCR from infected cells pretreated with SMER28 or rapamycin at 22 hpi. Mean ± SE for three independent experiments normalized to ETOH control; *p < 0.05, Student's t test.