Figure 1.

Blocking protein synthesis does not affect KCC2 protein level. A, Application of 100 μm cycloheximide (CHX) or 100 μm emetine inhibited the incorporation of [³⁵S]methionine in hippocampal slices (autoradiogram, top). SDS-PAGE demonstrated that similar amounts of protein were loaded (Coomassie-stained gel, bottom). B, Immunoblots with homogenates from hippocampal slices incubated in CHX and emetine. The tubulin signal (55 kDa) confirmed that similar protein amounts were loaded. The KCC2 signal (∼140 kDa) did not differ significantly from the corresponding controls (ctrl) after 4 h under arrested protein synthesis (ctrl: 90.3 ± 5.2%; CHX: 85.3 ± 8.4%; emetine: 90.6 ± 8.0%; p = 0.824). Statistical significance was assessed by Kruskal–Wallis one-way ANOVA on Ranks with Mann–Whitney's post hoc test. C, Application of 50 μm leupeptin (leu) for 4 h did not induce significant changes in the KCC2 protein level in hippocampal slices (105.1 ± 10.6%). Statistical significance was assessed by one-way ANOVA. The values for n are given in the bar diagram. Error bars denote SEM.