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. Author manuscript; available in PMC: 2015 Feb 5.
Published in final edited form as: Sci Signal. 2014 Aug 5;7(337):ra74. doi: 10.1126/scisignal.2005147

Fig 1. Itk association with CaM is Ca2+ dependent and enhanced by PI(3,4,5)P3 binding.

Fig 1

(A) CaM coprecipitation of endogenous Itk or Btk from mouse splenocyte lysates was perfomed with the indicated concentrations of Ca2+ in the precipitation buffer. Affinity purification with PI(3,4,5)P3-coated beads served as a positive control. (B) Direct comparison of recombinant Itk PH domain binding to PI(3,4,5)P3, Apo-CaM (EGTA), and Ca2+/CaM (Ca2+). (C) Effect of PIP3, IP4 and PIP2 addition on the coprecipitation of Itk or calcineuron with CaM from primary T cell lysates. (D) Dose-dependent enhancement of Itk binding to CaM by addition of PI(3,4,5)P3. All data are representative of 3 experiments. PIP2, PI(4,5)P2; PIP3, PI(3,4,5)P3; IP4, Ins(1,3,4,5)P4; ppt, precipitation; rItkPH, recombinant Itk PH domain