Fig. 5.

Modulation of P2X7R reduces the effect cytotoxic generated by ATP in high concentrations. a Pharmacological modulation of P2X7R was performed by cell counting after treatment with A740003 10 μM, a specific antagonist of P2X7R, suramin 30 μM, and PPADS 10 μM as unspecific antagonist of P2X receptors and ATP 5 mM. The treatment was performed after sequential reduction of FBS concentration, and the cell counting was performed after 48 h posttreatment. The difference in relation to control was determined as **p < 0.01 and ***p < 0.001. The difference of ATP 5 mM was determined as #p < 0.05 and ###p < 0,001. b Representative Western blotting assays showing positivity to P2X7R in KYSE450 GFP−/− and KYSE450 siP2X7R. c Relative P2X7R levels obtained by analysis of protein bands detected by Western blot were compared to GAPDH expression levels and the significance was described as **p < 0.01 and indicated difference in relation to GFP−/−. d Effect of P2X7R silencing on cell viability. After 48 h of silencing, the cells were plated and MTT assays were performed following 24 h. The significance was described as ***p < 0.001 and indicated difference in relation to GFP−/−. e Effect of ATP (2.5 and 5 mM) treatment on siP2X7R cell viability. The experiments were performed three times in triplicate