GST-moesin constructs were fused to glutathione-Sepharose beads as described previously (Bravo-Cordero et al., 2013 (link)). In brief, GST-moesin (pGEX4T1, tac promoter) was expressed in Escherichia coli and purified using glutathione-agarose beads. Talin fragments constituting the talin head domain (aa 1–433), talin rod R1–5 (aa 434–1,203), talin rod R6–10 (aa 1,205–1,971), or talin rod R11-DD (aa 1,975–2,541), R11 (aa 1,975–2,140), R12 (aa 2,141–2,291), and R13-DD (aa 2,300–2,541) were generated by PCR, excised with BamHI or SalI and NotI, and cloned into the pET30a vector (EMD Millipore). The resulting constructs were expressed in E. coli BL21, and protein expression was induced with 0.2 mM IPTG as described previously (Xing et al., 2001 (link)). Recombinant his-tagged talin protein was then purified using a HisTALON column according to the manufacturer’s instructions (Takara Bio Inc.). Protein concentration was determined and 1 µg of recombinant talin was added to 50 µl of GST-moesin beads for 2 h at 4°C, washed to remove unbound protein, and run on SDS-PAGE, as described previously (Lad et al., 2007 (link); Gingras et al., 2009 (link); Bouaouina et al., 2012 (link)).