Fig. 2

P2X7-mediated AMPK activation depends on mitoROS and Ca/CaMKK in BV-2 cells. BV-2 cells were pretreated with A438079 (10 μM) for 30 min, followed by the stimulation with BzATP (200 μM) for 15, 30 or 60 min. Then Fluo-3 AM (1 μM) (a) or MitoSOX (5 μM) (b) were used to determine cytosolic calcium and mitochondrial ROS, respectively. The fluorescence intensity was presented as percentages of the control group without any treatments. Data were the mean ± S.E.M. of at least 3 independent experiments. *p < 0.05, indicating the significant stimulating effect of BzATP; #p < 0.05, indicating the blockade effect of A438079. In c-f cells were pretreated with STO-609 (10 μM) (c) or mitoTEMPO (250 μM) (d-f) for 30 min, followed by the treatment with either BzATP (200 μM) or nigericin (10 μM) at the indicated time points. Cell lysates were used for immunoblotting (c, d and f) and in some experiments cytosolic calcium level was determined (e). Data were representative of 3 independent experiments. *p < 0.05, indicating the significant effects of BzATP; #p < 0.05, indicating the antagonist effect of A438079 and mitoTEMPO on the effects of BzATP. Data of a, b and e were the mean ± S.E.M. from 3 independent experiments. Data quantifications of c, d and f were shown in Additional file 2: Figure S2