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. Author manuscript; available in PMC: 2008 Jan 12.
Published in final edited form as: Cell. 2007 Jan 12;128(1):141–156. doi: 10.1016/j.cell.2006.11.040

Figure 4. K13 and K289 are ubiquitinated in vivo.

Figure 4

(A) PTEN shows adduct formation in vivo and a smear (poly) at 36 hours post transfection. Molecular weights in kD. ‘n’ denotes non-specific band and asterisks indicate migration of unmodified (single) and modified PTEN (double).

(B) Discrete adducts co-migrate with various PTEN-fusions. Asterisks as in (A). Molecular weights in kD.

(C) Immunoprecipitation of gfp-PTEN after HA-Ubiquitin co-expression confirms discrete mono- (labeled 1–3) and poly-Ub adducts (labeled poly). Migration of gfp-PTEN is shown. Molecular weights in kD.

(D) Low steady state levels (left panel), strong ubiquitination (middle panel) and PTEN-specific ubiquitination (right panel) of the gfp-PTEN-UbKØ fusion. Molecular weights in kD.

(E) Defects in mono-ubiquitination of the indicated lysine mutants. Left panel shows overview (location of stacking gel and molecular weights are indicated) and right panels show magnification and quantification of ratios for mono-Ub : main PTEN-band intensities.