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. 2012 Mar 23;13(5):455–461. doi: 10.1038/embor.2012.37

Figure 2.

Figure 2

c-Jun N-terminal kinase (JNK)-stimulated phosphorylation of BMAL1 and CLOCK proteins. (A,B) HEK293T cells were transfected with Myc–CLOCK/pSG5, BMAL1/pcDNA3.1 and JNK1/pSRα. Ser 520, Thr 527 and Ser 592 in mouse BMAL1 were mutated to Ala (mut3A). The cells were collected 30 min after the sorbitol stimulation (indicated concentrations) and were subjected to immunoblot analysis. Data are means with s.e.m. (n=3). Double asterisks indicate P<0.01 (Student's t-test, versus WT 0 mM). (C) NIH3T3 cells were transfected with Myc–CLOCK/pSG5, BMAL1/pcDNA3.1 and MKK7–JNKs/pCI. Total amount of DNA was adjusted by adding empty plasmids. The transfected cells were collected and subjected to immunoblot analysis. Lys 93 in JNK3 was mutated to Ala for a kinase-dead (KD) mutant, MKK7–JNK3(KD). WT, wild type.