Effect of ASC on DCs glycolytic pathway.

<p>To determine the influence of ASC on DC glycolysis DCs. A). DCs were stimulated with <i>C</i>. <i>muridarum</i> for 30 minutes and real-time ECAR was measured in a live DCs using an XFe-96 analyzer. Vertical lines indicate addition of glucose (glycolysis substrate), oligomycin (ATP synthase inhibitor), and 2-DG (glycolysis inhibitor). Graphs in this figure represent the mean ± SD of three independent experiments. B). Culture supernatants from WT and ASC^-/- DCs stimulated at a ratio of 1:5 with <i>C</i>. <i>muridarum</i> at 0, 1 and 2 hours were collected, and their pyruvate concentration in μM/ml was determined. Data were calculated as the mean ± SD for triplicate cultures from each experiment (*, P < 0.05). Control groups included uninfected DCs and culture media. n = 12 pooled from three independent experiments. In ASC^-/- DCs, the pyruvate production was markedly reduced compared to WT DCs after <i>Chlamydia</i> stimulation. C). Western blotting was used to analyze HEK II, ENO-1 and PDH in lysates from <i>C</i>. <i>muridarum</i> stimulated WT and ASC^-/- BMDCs after 0, 1 and 2 hours. The expression levels of the glycolytic enzymes HEK II and ENO-1 in WT and ASC^-/- DCs were unchanged before or after <i>C</i>. <i>muridarum</i> stimulation. PDH expression was downregulated during <i>C</i>. <i>muridarum</i> stimulation of ASC^-/- DCs.