Figure 1.

UBE3A expression in Mecp2-null mouse brain. (A) A multiple tissue microarray containing wt and knock-out mouse cerebral cortex samples (16) was stained for immunofluorescence using a C-terminal reactive anti-MeCP2 and an anti-UBE3A antibody. The MeCP2-negative cells (Mecp2 mt-expressing, blue histograms, percentage shown) were separately gated from MeCP2-positive cells (Mecp2 wt-expressing, red histograms) and the total population (black histograms) and compared with age-matched wt control female or male samples (orange histograms). (B) The graph shows combined LSC results (mean ± SEM) from four replicate slides using two different anti-UBE3A antibodies. (C) Protein extracts from whole adult mouse brain were probed on an immunoblot with anti-UBE3A or anti-GAPDH. A representative image shows lower expression in brain of UBE3A in both hemizygous male (−/y) and heterozygous female (−/+) brain compared with wild-type (wt, +/y and +/+) littermate controls for both Mecp2^tm1.1Bird and Mecp2^tm1.1Jae strains. The graph demonstrates combined GAPDH-normalized results (mean ± SEM, quantitation as previously described (32), example ratios shown below each lane) from eight different −/+ and six different +/+ brain samples from Mecp2^tm1.1Bird and Mecp2^tm1.1Jae strains. ^*P < 0.05, ^****P < 0.0001 by t-test.