Regulation of SREBP2 by mCMV infection and IFNβ treatment.

(A) Comparison of cleaved SREBP2 protein in mock-infected (lane 1), mCMV-infected (MOI of 1) (lane 2), mock-treated (lane 3), IFNβ- (50 U/ml) (lane 4), or IFNγ- (50 U/ml) treated (lane 5) BMDM for 24 h by Western blot analysis using YY1 as a loading control. Arrow indicates SREBP2 cleaved form that is induced upon lovastatin and ezetimibe treatment from liver extracts of cholesterol-fed mice (see Figure S8). The blot is representative of two independent experiments with biological triplicates for each experiment. (B) Wild type BMDM were infected with mCMV for 1 h. De novo RNA was labeled between 360 and 390 min post-infection, isolated, and hybridized to Affymetrix Gene 1.0 ST microarrays (Materials and Methods). After scanning and data capture, gene expression in mock-infected or infected cells was analyzed, and for the purposes of presentation, Srebf2 gene expression values from control (mock-infected) BMDM (black) were adjusted to a value of 1. Values for expression in infected cells (white) were then expressed as a number relative to the control. (C) BMDM from wild type or IFNAR1−/− knockout mice were treated with 10 U/ml of IFNβ or infected with mCMV. After 24 h, RNA was collected and the gene expression of Srebf2 was measured by qRT-PCR. Results show the level of gene expression of the treated or infected samples relative to the mock-treated samples. Bars represent the mean ± SD of biological quadruplicates. *pppt test.