Supplementary Figure 10: Impairment of dendritic localization of ITPR1 3' UTR by dominant-negative forms of DISC1.

(a) Inhibitory effect of hDISC1-N1-ΔARM on interaction between DISC1 and HZF. His pulldown assays were performed in the COS7 cell lysates expressing GFP-hDISC1-FL and the indicated Myc fusion proteins. 20 pmol of the purified recombinant His-HZF or His-GST protein was mixed with the cell lysates. Proteins that were bound to the immobilized beads with His fusion proteins were eluted and analyzed using immunoblot analyses with anti-GFP, anti-Myc and anti-His. Aliquots of original samples (1% input) and eluates (10%) in a were subjected to SDS-PAGE. (b) Dendritic localization of Venus–ITPR1 (IP3R1) 3' UTR in the neurons expressing hDISC1-N1-ΔARM. Cultured hippocampal neurons were transfected with the indicated plasmids at DIV 12. The transfected neurons were immunostained with antibodies to Venus (green) and MAP2 (red) at DIV 14. The blue bars indicate the distance between the soma and the farthest transported granules containing ITPR1 3' UTR. Magnified images of these granules are shown in the boxed areas. Scale bars, 15 μm.