JCS014423 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Localisation of GFP-OCRL1 in nocodazole-treated HeLa cells. HeLa cells expressing GFP-tagged OCRL1 treated with 5 μg/ml nocodazole for 2 hours before labelling with antibodies against ERGIC53, GM130 or TGN46. Bar, 10 μm. Insets show a magnified view of the boxed area.

• Supplemental Figure S2 -

  Fig. S2. Localisation of GFP-INPP5B in HeLa and NRK cells. (A) HeLa cells transiently expressing GFP-INPP5B (green) were fixed in paraformaldehyde (PFA) and labelled with antibodies against GM130 or ERGIC53, as indicated (red). Bar, 10 μm. (B) NRK cells transiently expressing GFP-tagged INPP5B (green) were fixed in methanol and labelled with antibodies against p115 (red). Bar, 10 μm. Insets show a magnified view of the boxed area.

• Supplemental Figure S3 -

  Fig. S3. Localisation of FLAG-tagged INPP5B in HeLa cells. HeLa cells transiently transfected with vector encoding FLAG-INNP5B (green) were incubated at 37°C (top) or 15°C for 3 hours (bottom), fixed in methanol and labelled with antibodies against GM130 or ERGIC53, as indicated (red). Bar, 10 μm. Asterisks indicate cells expressing FLAG-INPP5B with an altered ERGIC53 distribution.

• Supplemental Figure S4 -

  Fig. S4. Localisation of GFP-INPP5B D524A to the ERGIC and endosomes. HeLa cells were transiently transfected with vector encoding GFP-INPP5B D524A (green), fixed in methanol and labelled with antibodies to β-COP or the transferrin receptor (TfR), as indicated (red). Bar, 10 μm. Insets show a magnified view of the boxed area.