Fig. 3
KTR Technology Is Generalizable to Other Kinases (A) 3T3 cells expressing JNK KTR were stimulated with anisomycin (50 ng/ml), imaged, and quantified as described in the Experimental Procedures. Schematic representation of the engineered reporter is shown for all panels. Data represent the mean ± SD of more than 100 cells. Cells were preincubated with media (control), 10 ?M JNK inhibitor VIII (JNK inh.), 10 ?M SB203580 (p38 inh.), or 100 nM PD032591 (ERK inh.). (B) 3T3 cells expressing p38 KTR were stimulated with anisomycin (50 ng/ml), imaged, and quantified as described in the Experimental Procedures. Cells were pretreated or not with kinase inhibitors as in (A). (C) 3T3 cells expressing ERK KTR were stimulated with basic fibroblast growth factor-2 (bFGF2, 100 ng/ml), imaged, and quantified as described in the Experimental Procedures. Cells were pretreated or not with kinase inhibitors as in (A). (D) 3T3 cells expressing PKA KTR were stimulated with the PKA activator forskolin (10 ?M) imaged and quantified as described in the Experimental Procedures. Cells were pretreated with the specific PKA inhibitor H89 (30 ?M) (+ PKA inh.) or not (control). Where indicated, H89 (30 ?M) was added at time 120 min (+ PKA inh. at t = 120). |
Reprinted from Cell, 157, Regot, S., Hughey, J.J., Bajar, B.T., Carrasco, S., Covert, M.W., High-sensitivity measurements of multiple kinase activities in live single cells, 172417341724-34, Copyright (2014) with permission from Elsevier. Full text @ Cell