Fig. 3
Levels of phospho-Cx43 in Thy-1-stimulated astrocytes under proinflammatory conditions. A Kinetics of changes in pS373Cx43, Cx43, pS473AKT and AKT protein levels in Thy-1-Fc-stimulated primary rat astrocytes previously treated for 48 h with TNF (10 ng/ml). Hsp90 α/β was used as a loading control. B Quantification of the average values for the pS373Cx43/Cx43 ratio obtained by densitometric data analysis of the results shown in (A). Values are mean ± s.e.m. from 3 separate experiments and indicate the ratio between the densitometric values of the bands and the respective Hsp90 α/β control value. Astrocytes derived from rat brains, treated or not with TNF for 48 h (C, D). Astrocytes derived from hSOD1G93A or hSOD1WT mice (E, F) incubated for 30 min with either the vehicle (0.03% DMSO), the AKT inhibitor (AKTi, 3 μM), or the PI3K inhibitor LY294002 (LY, 3 μM). pS373Cx43 protein levels were evaluated by immunoblotting analysis of extracts from cells stimulated with Thy-1-Fc for 10 min or treated with TRAIL-R2-Fc as a control. D, F Quantification of the average pS373Cx43/Hsp90 α/β ratio values obtained by densitometric data analysis of the results shown in C and E, respectively. Values are mean ± s.e.m. from 3 independent experiments *p<0.05, **p<0.01 (ANOVA, Bonferroni post-test)