Fig. 8

Analysis of a putative second dimerization region in FLNc. a Yeast-two-hybrid experiments investigating interaction of FLNc Ig-like domains 15–23 (prey) with different fragments of FLNc as baits. Interaction was found with d15–23, d18–21 and d19–21 of FLNc. Note that deletion of the FLNc specific insertion in Ig-like domain 20 (d19–21Δ) abolished the interaction. +: interaction; −: no interaction. b FLNc d18–21 with and without (Δ) insertion in d20, and FLNa d18–21 were incubated in the presence of the cross linker EGS. Separation by PAGE and western blotting with an antibody against the immunotag of the recombinant fragments indicated oligomerization of FLNc d18–21 (arrow) but not of the other constructs. c GST pull down assay indicating that GST-FLNc d19–21 (GST + FLNc) binds T7-tagged FLNc d19–21, but not T7-tagged FLNc d15–17. GST alone does not bind any of the FLNc fragments. lc: loading control showing the correct size of the FLNc fragments