Figure 7. PTENα and PTEN form a complex and collaborate in energy metabolism.
(A) S-HA-tagged PTEN and FLAG-HA-tagged PTENα were transfected into 293T cells prior to S protein pull-down (S-PD). FLAG or HA immunoblotting was performed to detect PTEN-associated PTENα.
(B) In vivo binding of PTENα with PTEN. Endogenous PTENα was immunoprecipited using anti-αN antibody from mouse brain tissues for immunoblotting of PTEN.
(C) Evaluation of PINK1 expression in PTENα depleted cells by Western blotting.
(D) PINK1 expression was assessed in Pten-/- MEFs transfected with PTEN, PTENα, or PTEN+PTENα.
(E) Pten-/- MEFs transfected individually or in different combinations with PTEN, PTENα and PINK1, followed by analysis of ATP production. Data are presented as mean±SD. Labeling for statistical significance above each column indicates a comparison with the control column. n.s. not significant, p>0.05; *,p <0.05; **, p <0.01; ***, p <0.001.
(F) A graphic model of PTENα translation and its function in mitochrondrial energy metabolism. PTENα is synthesized through an eIF2A- and palindrome-dependent CUG initiation mechanism. PTENα forms a complex with canonical PTEN and these molecules collaborate in mitochondrial bioenergetics through regulation of cytochrome c oxidase activity and ATP production.
See also Figures S6 and S7.
