HCMV Targets the Metabolic Stress Response through Activation of AMPK Whose Activity Is Important for Viral Replication
Figure 2
Impact of AMPK inhibition on AMPK substrate accumulation during HCMV infection.
(A) MRC-5 human fibroblasts were incubated in glucose free media with DMSO (-) or AICAR for 1h or 2.5 h prior to blotting with antibodies specific for pSer79-specific ACC1, total ACC1, or tubulin. (B-D) Serum-starved MRC-5 human fibroblasts were mock infected or infected with HCMV (MOI = 3). After adsorption, cells were treated with the AMPK inhibitor, Compound C (5 µM), or DMSO (-). Cells were harvested at 24, 48 and 72 h post infection and analyzed by Western blot with antibodies specific for pSer79-specific ACC1, total ACC1 and tubulin in (B), TSC1, Glut4 and tubulin in (C), and pThr172-specific AMPK, total AMPK and tubulin (D). Relative pACC/ACC signal ratios in (B) were estimated during HCMV infection for DMSO and Compound C-treated cells using densitometry and subsequently normalized to the ratio of the DMSO control sample.