Supplementary Figure 1: BAR domain proteins binding to Endophilin.

a, Pull down using GST-SH3 domains for Endophilin A1, A2 or A3 and rat brain lysate. Interacting proteins were identified by mass spectrometry. Known interactors are shown in grey, members of the WAVE complex in black and BAR-domain proteins in blue. Asterisk denotes GST-SH3. List of identified proteins is provided in Supplementary Table 1. b, Pull-down using GST-SH3 domains of Endophilin A2 and cell extracts expressing EGFP-tagged indicated BAR proteins. Bin2 and OPHN1 were used as positive controls. GST was used as negative control. Binding proteins were detected by immunoblotting with an anti-EGFP antibody. ‘input’ lanes correspond to 1-5 % of the cell extracts. c, Schematic depicting the domain organization of the human full-length BAR domain-containing proteins cloned and used in this study. d, co-immunoprecipitation assays from cells co-expressing the indicated combination of EGFP- or Myc-tagged BAR domains from FBP17, CIP4 and TOCA-1 and showing that all three can heterodimerize. Input (‘I’, which corresponds to 10% of cell extracts), unbound (‘U’) and bound (‘B’) fractions for each were immunoblotted with anti-EGFP and anti-Myc antibodies. All experiments were repeated independently at least three times with similar results. Unprocessed original scans are provided in Supplementary Fig. 6.