Fig. 2

ADNP expression analysis of the cerebellum. (A) RT-PCR showing a significant increase in ADNP mRNA levels in the patient cerebellum compared to an age-and sex matched control subject (***p = 0.0001; unpaired student T-test). Gene expression values were normalized with three stable reference genes, i.e., β-Actin (BACT), β-2-Microglobulin (B2M), and Ubiquitin C (UBC). (B) ADNP protein expression analysis using an N-terminal antibody. Western blotting showed the presence of wild-type ADNP (150 kDa) in overexpression lysates with presence of the truncated protein. However, expression was absent in the patient as compared to the control, where wild-type ADNP could be visualized. (C) ADNP protein expression analysis using a C-terminal antibody. Western blotting showed the presence of wild-type ADNP (150 kDa) in overexpression lysates, as well as in the control subject, but not in the patient. GAPDH was used as a loading control for normalization