PTCHD1 protein, full-length human fetal brain PTCHD1 cDNA was PCR amplified and cloned into the pcDNA3.1/CT-GFP-TOPO expression vector (Invitrogen). After confirming the correct sequence and orientation of the insert, we transiently transfected COS-7 and SK-N-SH cells with 2 μg of purified construct DNA with SuperFect (Qiagen). 24 hours after transfection, we visualized the PTCHD1-GFP fusion protein in transfected cells using a Zeiss Axioplan 2 imaging microscope, equipped with the LSM510 array confocal laser scanning system, and the Zeiss LSM510 version 3.2 SP2 software package.
