JCS03487 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. The AP-1 complex does not affect the targeting of scNcr1p or scNpc2p. GFP-ALP, scNCR1-GFP and scNPC2-GFP plasmids were transformed into yeast deletions of the AP-1 complex (apl4Δ, apl2Δ, apm2Δ and aps1Δ). The GFP-tagged proteins were visualized by direct fluorescence microscopy. Corresponding DIC images are shown.

• Supplemental Figure S2 -

  Fig. S2. The AP-2 complex does not affect the targeting of scNcr1p or scNpc2p. GFP-ALP, scNCR1-GFP and scNPC2-GFP plasmids were transformed into yeast deletions of the AP-2 complex (apl3Δ, apm4Δ and aps2Δ). The GFP-tagged proteins were visualized by direct fluorescence microscopy. Corresponding DIC images are shown.

• Supplemental Figure S3 -

  Fig. S3. VPS4 does not direct the targeting of scNcr1p but plays a role in the trafficking of scNpc2p. scNCR1-GFP and scNPC2-GFP plasmids were transformed into yeast deletions of VPS4 (vps4Δ). The GFP-tagged proteins were visualized by direct fluorescence microscopy. Corresponding DIC images are shown.

• Supplemental Figure S4 -

  Fig. S4. Localization of mNPC1. AP-3^+/+ and AP-3^-/- mouse fibroblasts were infected with an adenovirus encoding mNPC1-GFP. The GFP-tagged protein (green) was assessed for co-localization with Vti1b and TfR (red) by indirect confocal microscopy using an anti-GFP antibody. Co-localization is visualized in yellow in the merged images. Enlarged images are magnified 400%. Bar, 10 μm.

• Supplemental Figure S5 -

  Fig. S5. Localization of LAMPI. The distribution of LAMPI (green) with Syn8 or Vti1b (red) was assessed by indirect confocal microscopy in AP-3^+/+ and AP-3^-/- mouse fibroblasts. Co-localization is visualized in yellow in the merged images. Enlarged images are magnified 400%. Bar, 10 μm.

• Supplemental Figure S6 -

  Fig. S6. Localization of hNPC2. AP-3^+/+ and AP-3^-/- mouse fibroblasts were infected with an adenovirus encoding hNPC2-GFP. The GFP-tagged protein (green) was assessed for co-distribution with Vti1b and TfR (red) by indirect confocal microscopy using an anti-GFP antibody. Co-localization is visualized in yellow in the merged images. Enlarged images are magnified 400%. Bar, 10 μm.