Figure 7.

Compensatory increases of DLG–MAGUKs levels in the Triton X-100-insoluble fraction of dissociated neuronal cultures. Subcellular fractionations with differential centrifugation and detergent extractions were performed on dissociated mouse or rat cortical cultures after DIV21, transduced at different time points with sh95 (C, GFP expression control; D0, transduction at DIV0; D7, transduction at DIV7; D12, transduction at DIV12). Ten micrograms of total protein were processed by SDS-PAGE and immunoblotted with antibodies directed against the indicated proteins (Mort, mortalin). A, Characterization of different fractions (H, homogenate; S2, cytosolic and microsomal fraction; P2, crude synaptosomes; 1T-P, Triton X-100 extraction pellet; 1T-S, Triton X-100 extraction supernatant). B, C, Sample immunoblots of the 1T-P fraction (top three) and P2 fraction (PSD-95) of indicated proteins is illustrated on the top and quantified protein amounts (mean ± SEM) normalized against the mean of control condition on each blot is plotted against the experimental condition [n = 6 rat (B) or mouse (C) cultures]. Statistical significance is indicated with asterisks, tested with ANOVA and post hoc analysis of experimental condition versus control or when indicated with bar among all indicated conditions. D, E, Sample immunoblots of mouse hippocampal cultures transduced with sh102 (D), sh97 (E), or control lentiviral vector, probed with indicated antibodies.