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. Author manuscript; available in PMC: 2022 May 9.
Published in final edited form as: Sci Signal. 2021 Nov 9;14(708):eabg2648. doi: 10.1126/scisignal.abg2648

Fig. 7. In vitro CLP257 treatment upregulated membrane KCC2 expression and Ser940 phosphorylation.

Fig. 7.

(A) KCC2, Ser940, and Thr1007 in the plasma membrane (M) and cytosol (C) for all treated P7 wild-type (WT) brain slices. (B) KCC2, Ser940, and Thr1007 in the membrane and (C) cytosol for all treatment groups plotted as percent of naïve. Number of WT P7 pups: n=8 (50 μM CLP257), n=7 (100 μM CLP257), n=4 (500 μM CLP257), n=3 (100μM CLP290). (D) Graphical representation of S940A+/+ knock-in mutant mice (41). (E to G) Western blotting for KCC2 expression in the plasma membrane and cytosol for all treated brain slices from S940A+/+ P7 pups (E). Quantified KCC2 expression in the membrane (F) and cytosol (G) for all S940A+/+ treatment groups plotted as percent of naïve. All proteins of interest in the cytosol were normalized to housekeeping protein β-actin; all proteins of interest in the plasma membrane were normalized to transferrin (TfR); and phosphoproteins were normalized to their respective total protein. Data plots show all data points with means ± SEM. * P<0.05 and ** P<0.01 by one-way ANOVA. S940A+/+ P7 pups: n=5 (50μM CLP257), n=5 (100μM CLP257), n=3 brain slices (500μM CLP257).