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. Author manuscript; available in PMC: 2016 Dec 17.
Published in final edited form as: Cell. 2015 Dec 10;163(7):1716–1729. doi: 10.1016/j.cell.2015.11.045

Figure 2. Genetic inhibition of the lipid biosynthetic program primes antiviral immunity.

Figure 2

(A) qPCR analysis of murine gammaherpesvirus-68 (MHV-68) genes in Control or SCAP−/− BMDMs infected with MHV-68 (MOI=0.5) for 48h. (B) qPCR analysis of HIV-1 mRNA (left) and representative immunoblots of HIV-1 p24 protein levels (right) from shControl or shSCAP THP1 macrophages 96h after infection. (C) MHV-68 titers from the lungs of LysM Cre+/− control (Control) or LysM Cre+/− SCAPfl/fl (SCAP−/−) mice on d.7 post intranasal infection (N=7; data shown is the combined results from two separate infection experiments of n=3 per group and n=4 per group). (D) qPCR analysis of indicated MHV-68 genes in WT BMDMs pretreated for 4h with Control or SCAP−/− conditioned media (C.M.) before MHV-68 infection (MOI=0.5) for 48h. C.M. from Control or SCAP−/− BMDMs was collected on d.7 post-differentiation. (E) qPCR analysis of Ifnb1 and representative interferon stimulated genes (ISGs) in unstimulated control or SCAP−/− BMDMs on d.8 of differentiation. (F) qPCR analysis of ISGs in ex vivo alveolar macrophage isolated from bronchoalveolar lavage from LysM Cre+/− control (Control) or LysM Cre+/− SCAPfl/fl (SCAP−/−) (3 mice/group). (G) qPCR analysis of indicated ISGs in unstimulated control or SCAP−/− BMDMs on d.9 of differentiation +/− 5ug/mL IFNAR blocking antibody for last 48h. (H) qPCR analysis of ISGs in WT BMDMs treated for 4h with Control or SCAP−/− conditioned media (C.M.). (I) MHV-68 ORF29 and ORF57 gene expression in WT BMDMs pretreated for 4h with Control or SCAP−/− conditioned media (CM) + 2ug/mL IFNAR blocking antibody before MHV-68 infection (MOI=0.5) for 48h. (J) qPCR analysis of Ifnb1 and Il1b expression in Control or SCAP−/− BMDMs unstimulated or stimulated with LPS (50ng/mL) or Poly:IC (1ug/mL) for 1h on d.8 of differentiation. All experiments are reported as means ± SD from three independent experiments, unless noted otherwise. *P < 0.05; **P < 0.01, ***P < 0.005 (two-tailed unpaired Student’s t test). See also Figure S3.