JCS012336 Supplementary Material

Files in this Data Supplement:

• Supplemental Table S1 -

  Adobe PDF

• Supplemental Table S2 -

  Adobe PDF

• Supplemental Figure S1 -

  Fig. S1. Screen for phenotypes caused by knockdown of candidate α-adaptin- and clathrin-interacting proteins. Quantification was performed on 3-4 experiments for each knockdown, with the value for each experiment averaged using 10-12 images (average integrated fluorescence per cell) and normalised using untreated cells in the same experiment (*P<0.05, ***P<0.005, t-test; error bars indicate s.e.m.).

• Supplemental Figure S2 -

  Fig. S2. Further examples of S2 cells showing Vps35-mRFP localization (magenta), double-labelled for endosomal markers (green), as in Fig. 3B.

• Supplemental Figure S3 -

  Fig. S3. Increased localisation to the plasma membrane (arrows) and leading edge of lamellopodia (arrowheads) of α-adaptin, dynamin, ubiquitylated proteins, EGFR, PVR, diphosopho-MAPK (ppMAPK) in primary haemocytes from vps35 third instar larvae, compared with the wild type. Note increased levels of ppMAPK in the nucleus (surrounded by broken line), panels are single confocal sections taken at 1 µm from the base of each cell.

• Movie 1 -

  Movie 1. A confocal stack of wild-type haemocytes stained for filamentous actin. Images were collected at 0.5-µm intervals starting from the top of the cells. Image brightness was substantially increased compared with Movie 2, to compensate for lower F-actin levels compared with vps35 cells. Images measure 53 µm square.

• Movie 2 -

  Movie 2. A confocal stack of vps35 haemocytes stained for filamentous actin. Images were collected as in Movie 1. Note the more pronounced lamellopodia compared with wild-type cells. These are partially detached from the surface of the coverslip and remain visible in confocal sections as far as 3 µm from the bottom of the cells. Images measure 53 µm square.