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. Author manuscript; available in PMC: 2015 Apr 29.
Published in final edited form as: BMB Rep. 2009 Mar 31;42(3):142–147. doi: 10.5483/bmbrep.2009.42.3.142

Fig. 1.

Fig. 1

Protein synthesis inhibitor-induced phosphorylation of CREB on Ser-108/111/114 requires CK1. (A), Diverse stress-stimuli induce the phosphorylation of CREB on Ser-108/111/114 in intact cells. HEK 293T cells were either mock-treated or exposed to cycloheximide (CHX, 20 μg/ml), UV light (20 Joules/m2), or ionizing radiation (IR, 5 Gy) and harvested at the indicated time intervals. Cell extracts were resolved by SDS-PAGE and immunoblotted with the indicated antibodies. (B), Anisomycin induces CREB Ser- 108/111/114 phosphorylation. HEK 293T cells were either mock- treated or subjected to 20 μg/ml anisomycin. Cell extracts were analyzed by immunoblotting with α-CREB and α-pCREB– S108/111/114 antibodies. (C), CHX induced CREB Ser-108/111/114 phosphorylation is sensitive to CK1 inhibition. HEK 293T cells were exposed to CHX with or without a 1 h-pretreatment with 75 μM D4476 (a CK1 inhibitor) for the indicated periods of time. The cells were then harvested and processed for immunoblotting with α-CREB and α-pCREB–S108/111/114.