Figure 3.
Catabolic Autophagy Was Not Required for the IFNγ-Mediated Suppression of MNV Replication
(A) Flow cytometry analysis of MNV replication (n = 3) at 12 hpi (MOI = 5) upon treatment with autophagy modulatory drugs. Chloroquine (CHQ, 20 μM), rapamycin (Rapa, 10 μM), wortmannin (WORT, 50 nM), or none (UN). Top: Representative flow data of MNV replication after no treatment (–IFNγ) or 100 U/ml of IFNγ pretreatment (+IFNγ) for 12 hr. Bottom: Quantitation of relative MNV replication after normalization to the level in untreated BMDMs. ∗ indicates statistically significant difference (p < 0.05) compared to the untreated (100%). This convention is carried through all figures. Mean virus titers ± SEM from three independent experiments (n = 3) are shown.
(B) Growth analysis of MNV (n = 3) in drug-treated BMDMs at 24 hpi (MOI = 0.05). n.s. indicates no statistically significant difference among samples (p > 0.05).
(C) Same analysis (n = 2) shown in (A) with more autophagy-modulating drugs: EBSS (Earle's balanced salt solution), BafilomycinA1 (100 nM), E64D/PepstatinA (10 μg/ml), LY294002 (10 μM).
(D) Same analysis (n = 2) shown in (B) with the drugs shown.
(E) Flow cytometry analysis of MNV replication (n = 3) upon the expression of dominant negative forms of autophagy proteins.
(F) Growth analysis of MNV (n = 3) in the transduced BMDMs.
(G) Flow cytometry analysis of MNV replication (n = 2) in control and Atg4BKO BMDMs.
(H) Growth analysis of MNV (n = 2) in control and Atg4BKO BMDMs.
In all graphs, data represent mean ± SEM. See also Figure S3.