Fig. 4

FSS influences pathways of energy metabolism and mitochondrial activity in trophoblasts. Metabolites of the glycolysis (a) and the TCA cycle (b) are presented for static and flow conditions, representing the peak area normalized to protein content and to the static control. Representative trace of reordered BeWo cells treated either under static (black) or flow (purple) pre-incubation expressing mtAT1.03. Dashed lines indicates the basal ratio used for further calculations. Red dotted lines indicate the deltas quantified (c). The maximum value after glucose removal was determined and the basal value subtracted to calculate the delta for hexokinase (d). Long-term glucose removal leads to decrease in the ratio (e). The minimum was determined before glucose re-addition and basal ratio was substracted. Oligomycin was added to the perfusion after the ratio reached a plateau phase after glucose re-addition. When the ratio reached the plateau under oligomycin the experiment was stopped (f),—either minimum or maximum was determined, depending on the cells reaction, and basal substracted (n = cells/days static 9/4; 0.95 dyne/cm² 7/4). Outlier were detected with Grubbs’ α = 0.05, normal distribution of data was analyzed with Shapiro–Wilk test. Statistical analysis for the relative abundance of metabolites was performed with a multiple unpaired t-test. Statistical analysis for the mitochondrial assay was performed using an unpaired t-test. Statistical significance was set at p < 0.05. Values represent mean ± SEM based on experiments using three to five different cell passages