Fig. 1.

ACLY is not essential for HCMV-induced lipogenesis. (A) Diagram of the synthesis of cytosolic Ac-CoA in mammalian cells; see text for details. (B) Protein levels of ACLY and ACSS2 in control HFs (CTRL) expressing a control sgRNA-targeting luciferase gene and the C9 HF line in which ACLY was knocked out. (C) ACLY knockout had little effect on HCMV growth in HFs. Confluent control and ACLY-KO C9 HFs were serum-starved and infected by HCMV at an MOI of 3. HCMV viral titers were examined at 72 and 96 hpi. (D) ACLY knockout reduced de novo lipid synthesis from glucose carbon by 50%. ACLY-KO cells and control HFs were mock-infected or were infected by HCMV at an MOI of 3. At 48 h, uninfected and infected cells were labeled with [U-¹⁴C]-d-glucose for 24 h; total lipids were extracted at 72 hpi and were counted by scintillation counter. Data are shown as the mean ± SD of triplicates. See details in Materials and Methods. (E) ACLY knockout had little effect on total cellular lipid levels in both mock- and HCMV-infected cells as measured by lipid droplet staining using BODIPY 558/568 C₁₂.