FIGURE 5.

Printor is found in both cytosolic and membrane-associated fractions. A, post-nuclear supernatant (PN) from SH-SY5Y cells was separated into cytosol (C) and membrane (M) fractions. Aliquots representing an equal percentage of each fraction were analyzed by immunoblotting with anti-printor, anti-torsinA, and anti-calnexin antibodies. B, the level of the indicated proteins in each fraction was quantified using NIH Scion Image and shown as a percentage of the total level of the indicated protein. Data represent mean ± S.E. from at least three independent experiments. C, SH-SY5Y cells expressing Myc-tagged printor were immunostained with anti-Myc and anti-KDEL, anti-EEA1, anti-LAMP2, or anti-TIM23 primary antibodies followed by detection with secondary antibodies conjugated to Texas Red (marker proteins, red) or FITC (printor, green). Hoechst stain was used to visualize the nucleus. Scale bars, 10 μm.