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. Author manuscript; available in PMC: 2008 Jul 11.
Published in final edited form as: Mol Cell. 2008 May 23;30(4):415–425. doi: 10.1016/j.molcel.2008.03.025

Figure 4. Phosphorylation of Bim on Ser65 and Thr112 in vivo.

Figure 4

A) Characterization of an antibody to the Bim phosphorylation site Thr112 on BimEL (Thr56 on BimL). 293T cells were transfected with an empty expression vector or a vector that expresses BimL. The expression of BimL and phosphoThr56-BimL was detected by immunoblot analysis of cells extracts. The effect of co-expression of constitutively activated JNK1 (JNK1*) and the replacement of Thr56 with an Ala residue is shown.

B) Homozygous BimLoxP, BimT112A, and Bim3SA primary MEF were serum-starved or treated with serum (S) (10% fetal bovine serum; 30 mins) or UV light (60 J/m2; 60 mins). Cell extracts were examined using antibodies to Bim, phospho-Thr112-Bim, phosphoSer65-Bim, and Tubulin.

C) Wild-type and Jnk1−/− Jnk2−/− (Jnk−/−) fibroblasts were serum-starved or treated with serum (S) or UV light. Cell extracts were examined using antibodies to Bim, phospho- Thr112-Bim, phosphoSer65-Bim, and Tubulin. The basal phosphorylation of BimEL on Ser65 was slightly increased in JNK-deficient fibroblasts compared with wild-type fibroblasts.

D) Jnk1−/− Jnk2−/− (Jnk−/−) fibroblasts were serum-starved or treated with serum and/or the MEK inhibitor U0126 (30 mins). Cell extracts were examined using antibodies to phospho-ERK, ERK, phospho-Thr112-Bim, and Bim.