Figure 3
From: Optogenetically controlled RAF to characterize BRAF and CRAF protein kinase inhibitors
Effect of kinase inhibitors on light-induced BRAF and CRAF signaling.
(A,C) HEK 293T cells expressing BRAF-CRY2 (A) or CRAF-CRY2 (C) were treated with the kinase inhibitors indicated for 26 h prior to further incubation in darkness (−) or illumination with blue light (+) for 5 min. Cell lysates were immunoblotted to detect BRAF, CRAF, pMEK and pERK as indicated. (B,D) SEAP activity was determined from supernatants of cells expressing BRAF-CRY2 (B) and CRAF-CRY2 (D) and treated with inhibitors as indicated for 2 h before further incubation in darkness (dark columns) or exposure to blue light (blue columns) for 24 h. Inhibitor concentrations used: dabrafenib 10 μM, vemurafenib 3 μM, GW5074 5 μM, trametinib 0.05 μM.
