Figure 1

Targeting pHSP20-14-3-3 protein interactions. A. A representative Western blot (n = 3 separate experiments) using antibodies to HSP20 (lane 1), phosphorylated cofilin (lane 2), and cofilin (lane 3). B. A representative SPR-based evaluation of HSP20 binding to a class of 14-3-3 proteins. Synthesized peptides containing a partial sequence of phosphorylated HSP20 were immobilized via amine-coupling to a BIAcore chip, and GST-HIS-14-3-3 isoforms (YWHAG, γ; YWHAH, η; YWHAZ, ζ; YWHAB, β; and YWHAE, ε) were injected at 20 μg/ml. Experiments were conducted in triplicate. C. A schematic drawing of the principle behind the fluorescence polarization (FP) assay. FP signals of a flourophore is defined here as, FP = (V-H)/(V+H); where V is the vertical component and H is the horizontal component of the emitted light when excited by vertical plane polarized light. D. Changes in FP signals in response to a number of compounds belonging to the PRLX24905 scaffold (USA Patent & Trademark, Publication 20090136561: "Non-peptidyl agents with pHSP20-like activity, and uses thereof"). Data are presented as mean ± SE (n = 3 separate experiments).