Supplementary Figure 5: NMDAR activation induces Panx1 phosphorylation at tyrosine 308.

a) Overlay and intensity line-scan of pY308Panx1 and Myc signals identifying the higher molecule weight species of Panx1 as the preferentially phosphorylated form. b) Phosphatase treatment of NMDA:Gly stimulated N2a cells expressing Panx1^WT. Cell lysates were treated with or without lambda phosphatase for 1 hour at 60°C to dephosphorylate Panx1. c) Inhibition of Panx1 phosphorylation at Y308 by the SFK inhibitor PP2 (10µM) in cells co-expressing Panx1^WT and Src^Y530F. *=p<0.05 compared to non treated cells (lane 1). d) Western blot analysis of Panx1 phosphorylation at Y308 in N2a cells co-expressing Panx1^WT or Panx1^Y308F and the kinase dead Src^K298M or constitutively active Src^Y530F. ** and ***=p<0.01 and 0.005 respectively. e) Immunoprecipitation efficiency for GluN1 in hippocampal slices. Pre=whole cell lysate before IP; Post=remaining fraction of GluN1 in unbound lysate (flow through) following IP. Full length blots are presented in supplementary figure 10.