Figure 4

Expression and localization of PTCHD3 isoforms. A: A multiple tissue northern blot demonstrating the expression profile of PTCHD3 isoforms in different tissues. B: A schematic demonstration of two predominant isoforms of PTCHD3 showing truncation of the last exon of previously documented PTCHD3 isoform (coding sequence: 2,304 bps) in the newly identified isoform (coding sequence: 1,602 bps). C: Cloning and expression of PTCHD3 isoforms tagged with a Myc epitope in COS-7 cells resulted in detection (by western blot) of two proteins of ~115 KDa (lanes 1 and 2) and ~80 KDa (lanes 3 and 4); lane 5: run with the lysates prepared from untransfected cells. D: Both PTCHD3 isoforms are localised to the endoplasmic reticulum (ER). Immunostaining of COS-7 cells transfected with constructs expressing Myc-tagged short or longer isoforms of PTCHD3 using antibodies detecting calnexin (red) as an ER marker and Myc epitope (green) demonstrated co-localization of PTCHD3 isoforms with calnexin to the ER (Pearson coefficient 0.85).