Inhibition of AMPK blocks HCMV-induced glycolytic activation.

<p>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. (<b>A</b>) At 48 h post infection, cells were labeled with [¹⁴C]-deoxyglucose for 5 min and harvested for scintillation counting as described in the <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002502#s4" target="_blank">materials and methods</a>. (<b>B</b>) At 48 h post infection, cells were labeled for 1 min with [¹³C] glucose containing DMEM, quenched with cold methanol, and processed for LC-MS/MS to measure the accumulation of [¹³C] fructose bisphosphate. All values are means + SE (n = 2). (<b>C</b>) At 40 h post infection, fresh media was added to all cells. At 58 h post infection, media was removed from mock and HCMV-infected fibroblasts and lactate secretion into the media over the 18 hr time frame was analyzed. All values are means + SE (n = 6). (<b>D</b>) Serum-starved MRC-5 human fibroblasts were HCMV infected (MOI = 3) and Compound C (5 µM) was added either immediately after adsorption (0) or 24 h post infection. Lactate excretion was measured from 40–58 hpi or from 58–76 hpi. Values are shown as the percent of the HCMV-infected, DMSO treated control and are means + SE (n≥3).