Identification of Small Molecules that Disrupt Signaling between ABL and Its Positive Regulator RIN1
Fig 2
Validation of TR-FRET screening assay for detection of RIN1::ABL binding.
Binding was quantified as a ratio of GFP emission at 520 nm to terbium emission at 486 nm. The negative control was normalized to 1. Experiments were performed in quadruplicate. (A) ATP was omitted from the buffer mix to prevent RIN1 phosphorylation by ABL. *p = 3.0x10-6 (B) 10 μM or 100 μM imatinib was added to inhibit ABL kinase activity. *p = 0.001 (C) Added untagged ABL competed with ABL-eGFP (1:1 and 10:1 ratios were used). *p = 1.4x10-7