Fig. 3

ENO1 knockdown DCs have altered metabolism and mitochondrial morphology. a & b Pyruvate Concentration in Chlamydia-pulsed Dendritic Cells. Pyruvate concentration was obtained in DC lysates and supernatant. Pyruvate concentration was measured using a spectrophotometer. Pyruvate concentration was determined after pulsing with Chlamydia at different time points for 0, 0.5, 1, 2 and 4 h. The concentration of pyruvate was significantly higher in WT cells compared to ENO1 knockdown DCs (p < 0.05). * Denotes statistical significance. This experiment was repeated 3 times. c Pyruvate dehydrogenase expression in Chlamydia-pulsed WT and ENO1 knockdown DCs. The total protein was detected using Cy5 dye and the protein of interest, in this case pyruvate dehydrogenase, was probed using Cy3 secondary antibody from the Amersham System (GE Healthcare, Pittsburgh, PA). Proteins were normalized using ImageQuant Reader (GE Healthcare​, Pittsburgh, PA). The ratio of Cy3/Cy5 indicates the differentially expressed proteins. This experiment was repeated 3 times. d TEM analysis of the WT and ENO1 knockdown DCs. The results indicate that there are morphological differences in the mitochondria of Chlamydia pulsed and nonpulsed WT and ENO1 knockdown DCs using TEM. This depicts the remodeling of the mitochondria to tubular and fused, with tight cristae in the WT DCs and to more circular, swollen and fissed with loose cristae in ENO1 knockdown DCs. e & f Expression of mitochondrial associated proteins. The blots shown are proteins associated with mitochondrial function. ANT 1/2/3/4 and TOM 20 were differentially expressed in Chlamydia pulsed ENO1 knockdown DCs. * denotes statistical significance. This experiment was repeated 3 times