Skip to main content
. Author manuscript; available in PMC: 2014 Jan 4.
Published in final edited form as: FEBS Lett. 2012 Nov 21;587(1):60–66. doi: 10.1016/j.febslet.2012.10.051

Fig. 1.

Fig. 1

Dab1 negatively regulates Cin85 phosphorylation in a RXXS/T motif. (A) The negative regulation of Cin85 RXXS/T phosphorylation by Dab1 is independent of Dab1 tyrosine phosphorylation. HEK 293 cells were transfected with the indicated constructs, starved of serum and treated with AP20187 (200 nM) for 30 minutes prior to lysis. Clarified extracts were either directly subjected to SDS-PAGE and immunoblotting with the indicated antibodies or first to anti-Flag immunopreciptation as indicated. (B) HEK 293 cells were transfected with Flag-Cin85, starved of serum and treated with or without 100 nM Calyculin A for 30 minutes prior to lysis as indicated. Clarified extracts were treated as in A. (C) HEK 293 cells were transfected with the indicated constructs. Co-transfected 10 cm dishes received 15 μg of Flag-Cin85 and either zero, 5 μg or 10 μg (as in A) of FKBP-Dab1. (D) Flag-Cin85 was immunoprecipitated from transfected cells, subjected to SDS-PAGE and stained with coomassie blue (upper panel). Whole cell extract or 10% of the immune complexes were subjected to immunoblotting with the indicated antibodies.