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. Author manuscript; available in PMC: 2015 May 6.
Published in final edited form as: Cell Metab. 2014 Apr 24;19(5):836–848. doi: 10.1016/j.cmet.2014.03.023

Figure 4. PTENα is localized predominantly in cytoplasm and mitochondria.

Figure 4

(A) Pten-/- MEFs transfected with N-terminal or C-terminal GFP-tagged PTEN or PTENα were subjected to protease protection assay, confirming the difference in subcellular distribution patters of PTENα and PTEN.

(B) Subcellular localization of C-terminal GFP-tagged PTENα (with an ATG>ATA mutation) and PTEN shown by confocal fluorescence microscopy. MitoTracker was used to indicate mitochondria. Overlay, merged images of GFP and MitoTracker.

(C) Cell fractionation was performed to isolate mitochondria in Pten+/+ and Pten-/- MEFs prior to immunoblotting analysis of PTENα and PTEN expression. W, whole cell lysate; M, mitochondria; C, cytoplasm. Cytochrome c and a-tubulin were used as mitochondrial and cytoplasmic markers.

(D) Mitochondria isolated from mouse brain cortex were subjected to sub fractionation of mitochondria prior to evaluation of PTENα by immunoblotting. Tom40 and Cox1 were used as markers for the mitochondrial outer membrane and inner membrane respectively. Cytochrome C is a dynamic component of mitochondria and can be found in both the inner membrane and intermembrane space.

See also Figures S4-S7.