Supplementary Materials

Supplementary Materials for:

Vitamin E Facilitates the Inactivation of the Kinase Akt by the Phosphatase PHLPP1

Po-Hsien Huang, Hsiao-Ching Chuang, Chih-Chien Chou, Huiling Wang, Su-Lin Lee, Hsiao-Ching Yang, Hao-Chieh Chiu, Naval Kapuriya, Dasheng Wang, Samuel K. Kulp, Ching-Shih Chen*

*Corresponding author. E-mail: chen.844{at}osu.edu

This PDF file includes:

  • Fig. S1. Effects of α-tocopherol and γ-tocopherol on normal prostate epithelial cell viability and the phosphorylation and intracellular localization of Akt in prostate cancer cells.
  • Fig. S2. Evidence that tocopherols promote dephosphorylation of Ser473 in Akt by recruitment of Akt and PHLPP1 to the plasma membrane.
  • Fig. S3. Effect of α-VE5 and γ-VE5 on the phosphorylation of Akt in prostate cancer cells.
  • Fig. S4. Evidence that α-VE5 and γ-VE5 promote dephosphorylation of Ser473 in Akt by recruitment of Akt and PHLPP1 to the plasma membrane.
  • Fig. S5. α-VE5– and γ-VE5–induced colocalization of Akt and PHLPP1 to the nonraft domains of the plasma membrane, essential role of PHLPP1 in α-VE5– and γ-VE5–induced Akt dephosphorylation, and in vivo tumor-suppressive activity of γ-VE5 in the LNCaP-abl xenograft tumor model.
  • Fig. S6. Membrane recruitment of Akt and PHLPP1 by tocopherols and VE5 compounds is PH domain–dependent.
  • Fig. S7. Immunocytochemical analysis of the time- and concentration-dependent changes in plasma membrane localization of PDK1 in γ-VE5–treated LNCaP cells.
  • Fig. S8. Peptide sequences and main secondary structural elements of the PH domains of Akt, PDK1, BTK, and ILK.
  • Table S1. Hematology values and body weight data from athymic nude mice after 21 days of treatment with γ-VE5.
  • Table S2. Serum chemistry values from athymic nude mice after 21 days of treatment with γ-VE5.
  • Table S3. SPR spectroscopic analysis of the binding affinities of α-VE5 and γ-VE5 for wild-type and mutated PH domains of various proteins.
  • Table S4. Sequences of primers.

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Citation: P.-H. Huang, H.-C. Chuang, C.-C. Chou, H. Wang, S.-L. Lee, H.-C.Yang, H.-C. Chiu, N. Kapuriya, D. Wang, S. K. Kulp, C.-S. Chen, Vitamin E Facilitates the Inactivation of the Kinase Akt by the Phosphatase PHLPP1. Sci. Signal. 6, ra19 (2013).

© 2013 American Association for the Advancement of Science