Fig. 4. Knocking down of purinergic receptors diminished the stimulatory effect of ATP. siRNA against P2RX and P2RY was individually transfected into hTCEpi cells using Lipofectamine 2000 reagent. Cell migration was recorded in the presence of 100 nM ATP at the EF strength of 30 mV/mm, and directedness (A) and migration speed (C) was calculated. n=50 cells for each group. Results confirmed in two separate experiments. Statistical analysis was performed. Data represented as mean SEM. n=50 for each group. Statistical analyses were performed between each group, for directedness (B) and for migration speed (D). *p<0.05, **p< 0.01, and NS not significant between two groups. (E and F) Apyrase treatment attenuated the effect of ATP. siRNA against P2X and P2Y was individually transfected into hTCEpi cells by using Lipofectamine 2000 reagent. Cell migration was recorded in the presence or absence of 100 nM ATP and 10 U/ml apyrase at the EF strength of 30 mV/mm, and directedness (E) and migration speed (F) was calculated. n=40-50 cells for each group. Statistical analysis was performed. Data represented as mean SEM. *p<0.05, and **p< 0.01 between two groups indicated in the Fig.. #p<0.01 vs. Control siRNA no ATP group (leftmost column). p<0.01 vs. Control siRNA 100 nM ATP group (second column from the left). p<0.01 vs. Control siRNA ATP+apyrase group (third column from the left).