Saturated Very Long Chain Fatty Acids Are Required for the Production of Infectious Human Cytomegalovirus Progeny
Figure 2
Identification of genes involved in fatty acid metabolism that are induced during HCMV infection.
(A) RT-qPCR analysis of cellular RNAs. RNA was analyzed at 48 hpi with BADinUL99GFP (10 IU/cell). Results were normalized to the expression level of 4 housekeeping genes, except in the case of elongases where GAPDH alone was used, and presented as log2-transformed fold change (infected/mock). Fold increases of ELOVL7, ACSL6, ELOVL3 and SLC27A2 are indicated. The results show the average of two independent experiments. (B) RT-qPCR analysis of acyl-CoA synthetase RNAs in HCMV-infected cells. Results are from data in panel A. Error bars represent ±1 SD of the mean. (C) ACSL1 protein accumulates during HCMV infection. Fibroblasts were mock-infected or infected with BADinUL99GFP (3 IU/cell), harvested after indicated time intervals and analyzed by western blot by using antibody to ACSL1. β-actin was monitored as a loading control. Band intensities were quantified using ImageJ software and ratio of ACSL1/β-actin after infection was compared to that of mock infected cells (M). (D) RT-qPCR analysis of elongase RNAs. RNA was analyzed at 48 hpi with BADinUL99GFP (10 IU/cell). Results are from two independent experiments assayed in triplicate. Error bars represent ±1 SD of the mean. (E) ELOVL3 protein accumulates in HCMV-infected cells. Cells were treated as in panel C and analyzed by western blot by using an antibody to ELOVL3. β-actin was monitored as a loading control. Band intensities were quantified as in panel C.