Fig. 6.

Acetate production and lipid synthesis from glucose in HFs depleted of PDH. (A) Depletion of PDH E1α in mock- and HCMV-infected HFs using two different shRNAs, shE1α #A and #B. (B and C) Depletion of PDH increased the accumulation of pyruvate and acetate derived from glucose in mock- and HCMV-infected HFs. HFs were treated with shGFP (control) or shE1α for 3 d, followed by serum starvation for 1 d. Cells then were mock or HCMV infected in serum-free DMEM. At 24 hpi the cells were labeled with 5.6 mM [1, 6-¹³C2]-d-glucose in serum-free DMEM. At 48 hpi the medium was collected for NMR quantitation of [3-¹³C]-pyruvate (B) and [2-¹³C]-acetate (C). Data are shown as the mean ± SD of triplicates. (D) Lipid synthesis from glucose carbon in mock- and HCMV-infected HFs depleted of PDH E1α. HFs were treated with shGFP or shE1α as described, and lipid synthesis from [U-¹⁴C]-d-glucose was measured in mock- and HCMV-infected cells as described in Fig. 1D. Data are shown as the mean ± SD of triplicates. (E) HCMV viral titers at 72 and 96 hpi in HFs treated with shGFP or shE1α as described in A.