Fig. 4.

P2X7 activation mediates SOD1^G93A release from NSC-34 motor neurons. A, B SOD1^G93A-EGFP (SOD1^G93A)-transfected NSC-34 cells, or C non-EGFP SOD1^G85R-, non-EGFP SOD1^G127X- or non-transfected (SOD1) NSC-34 cells were pre-incubated in the A, C absence or B presence of 10 μM AZ10606120 (AZ10) for 1 h. Cells were then incubated in the absence (0 mM or basal as indicated) or presence of A–C 1, 3 or A–C 5 mM ATP for 20 min and pelletable fractions obtained by centrifugation. A, B The presence of SOD1 in pelletable fractions was assessed by immunoblotting using an anti-EGFP antibody and densitometry. One representative immunoblot (upper image) is shown, and data (lower image) are presented as mean ± SD (n = 3) relative to A maximum release or B release in the absence of AZ10 and ATP; **p < 0.02, ***p < 0.01 compared to 0 mM ATP or corresponding basal, ^††p < 0.02 compared to ATP in the absence of AZ10. C The presence of SOD1 in pelletable fractions was assessed by immunoblotting using an anti-SOD1 antibody. Data are representative of n = 2–3 experiments