Fig. 5.

Alterations in Cav-1 and MTC-4 expression, mitochondrial volume, MMP, ATP, l-lactate, and ROS generation induced in fibroblasts by co-cultured with epithelial cells. a Representative images for detection of Cav1, MCT-4, and corresponding GAPDH by western blot. b, c Quantification of western blots for CAV-1 and MCT-4 expression showing significant decrease CAV-1 and up-regulation of MCT-4 in NOF when co-cultured with epithelial cells at distance, in transwells. Flow cytometric analysis of mitochondrial volume (d) and specific MMP (e) for fibroblasts alone or in epithelial–stromal units, showing a similar MTG and trendency of MMP drop after co-cultivation. f Luminescent analysis for ATP generation showing increased ATP production in NOFs co-cultured with NOKs compared to NOFs alone and decreased ATP level in NOFs seeded with OSCCs. g, h Measurement of l-lactate generation and flow cytometric analysis of ROS production showing gradually upregulated l-lactate production and ROS level from NOFs alone, to NOFs co-cultured with NOKs, and NOFs co-cultured with OSCCs. The data points were presented from six different patient sets in d–f. The data points were presented from the replicates on the averages of the six different patient combinations in g and h. All points represent mean values and SEM. Independent experiments were performed more than three times. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; Wilcoxon signed-rank test was used in d–f; paired t test was used in b, c, g and h