Figure 2
Change of KCC2 expression involved in the progress of epileptiform activity in hippocampal slices. (A) Extracellular field potential recording of epileptiform activities in DG granule cell layer induced by 50 μM CTZ. Sample traces showing baseline (upper), interictal-like spike discharge (middle) or HFHA bursting activity (bottom) at different time points after CTZ treatment. (B) Dot plot showing the individual and the group data of the latency of interictal-like spike or ictal-like bursting activities after CTZ treatment. (C) Immunostaining images showing the downregulation of KCC2 in DG after CTZ treatment for 2 hr. Scale bar, 20 μm. (D,E) Western blot results showing the change of membrane KCC2 expression at 0.5, 1 and 2 hr (D), or membrane pS940 KCC2 at 1 and 2 hr (E) after CTZ or DMSO treatment, respectively. The gels were run under the same experimental conditions and were cropped around 140 and 46 KDa. The full-length gels were presented in Supplementary Figs 3 and 5. Bar histogram showing the quantification of KCC2 expression or value of membrane pS940/total membrane KCC2 normalized to control. Membrane KCC2 or pS940/membrane KCC2 level was significantly downregulated at 1 hr (*P < 0.05, **P < 0.01) or 2 hr time point (**P < 0.01). (F) Sample western blot showing membrane NKCC1 expression was low and no significant change at 1 and 2 hr after CTZ treatment. The gels were run under the same experimental conditions and were cropped around 140 and 46 KDa. The full-length gels were presented in Supplementary Fig. 6. Bar histogram showing the quantification of NKCC1 expression normalized to control.
