JCS02930 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 5 (Adobe PDF) -

  Fig. S5. Primers and PCR conditions used to generate plasmid constructs utilized in this study

• Supplemental Figure 1 -

  Fig. S1. Colocalization between endogenous OA1 and various markers by confocal analysis in MNT1 cells. (A) Examples of colocalization between OA1 and various markers, as visualized by yellow/orange areas, which predominate with LAMP1 and Pmel17. (B) An example of how percent of colocalization was evaluated. In the cell shown here, out of 25 vesicles positive for OA1, 21 were also positive for Pmel17 (84%). Immunofluorescence conditions are as in Fig. 1B. Bars, 10 mm.

• Supplemental Figure 2 -

  Fig. S2. Subcellular distribution of LAMP/OA1 chimeras by confocal analysis in MNT1 cells. (A) An example of colocalization between LAMP/i3DFY and farnesylated GFP, which labels the plasma membrane. (B) Examples of colocalization between LAMP/i3FY or LAMP/CT2M5 and endogenous OA1, as visualized by yellow/orange areas, which are more extensive with LAMP/CT2M5. (C) An example of how percent of colocalization was evaluated. In the cell shown here, out of 25 vesicles positive for LAMP/CT2, 23 were also positive for endogenous OA1 (92%). Immunofluorescence conditions are as in Fig. 5. VE, vesicular pattern; PM, plasma membrane pattern. Bars, 10 mm.

• Supplemental Figure 3 -

  Fig. S3. Colocalization between OA1CT2-mycHis and various markers by confocal analysis in MNT1 cells. Immunofluorescence conditions are as in Fig. 9A. Yellow/orange indicates areas of colocalization, which are mostly seen with LAMP1 and Pmel17. L.Y., Lucifer Yellow at 1 hour chase. Bars, 10 mm.

• Supplemental Figure 4 -

  Fig. S4. Colocalization between OA1CT2-mycHis single mutants and endogenous OA1 by confocal analysis in MNT1 cells. Yellow/orange indicates areas of colocalization with endogenous OA1, which are less extensive with OA1CT2 WE®AA. Immunofluorescence conditions are as in Fig. 10C. Bar, 10 mm.