(A) Free cholesterol distribution in wild type and NPC2 (−/−) fibroblasts.
(B) PV replicon replication in wild type and NPC2 (−/−) fibroblasts. Mean data +/−SEM with 6 replicates each is plotted.
(C) Peak PV replication levels for wild type, NPC1 (−/−) and NPC2 (−/−) fibroblasts. Mean data +/− SEM from 5 independent experiments, normalized by the expression of reporter luciferase RNA, is plotted. ***: p<0.001
(D) Cholesterol and viral dsRNA distribution in wild type and NPC2 (−/−) fibroblasts.
(E) Free cholesterol levels within wt and NPC2 (−/−) fibroblast replication organelles. Mean data +/−SEM, from wild type (n=30) and NPC2 (−/−) (n=30) fibroblasts plotted.
(F) Acute cholesterol extraction inhibits PV replicon replication in NPC2−/− fibroblasts. Mean data +/− SEM from 2 independent experiments is plotted.
(G) 3CDpro / 3Dpol ratio in wild type and NPC2 (−/−) fibroblasts. Mean data +/−SEM from 3 independent experiments are plotted.
Scale bars 10µm.
Figure 4 related to Figure S4.