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. Author manuscript; available in PMC: 2016 Jul 4.
Published in final edited form as: Nat Commun. 2011 Jun 28;2:371. doi: 10.1038/ncomms1372

Figure 5. Phosphorylation of T371 protects TRF1 from being targeted for degradation.

Figure 5

(a) Cycloheximide chase experiments. Nocodazole-arrested HeLaI.2.11 cells were treated with cycloheximide (100 μg/ml) in the presence of 1 μM okadaic acid for the indicated times. Okadaic acid was used to prevent dephosphorylation of T371. Immunoblotting was performed with anti-TRF1, anti-pT371 or anti-γ-tubulin antibody. (b) Quantification of total TRF1 and phosphorylated (pT371)TRF1 from (a). The signals from the western blots were quantified with densitometry. The level of total TRF1 or phosphorylated (pT371)TRF1 is represented in arbitrary units after their signals were normalized relative to those of γ-tubulin. Standard deviations from three independent experiments are indicated. (c) Cycloheximide chase experiments. HT1080 cells expressing Myc-TRF1, Myc-TRF1-T371A or Myc-TRF1-T371D were treated with 100 μg/ml cycloheximide for the indicated times, followed by immunoblotting of the lysates with anti-Myc or anti-γ-tubulin antibody. (d) Quantification of Myc-tagged wild type TRF1 (blue bars), Myc-tagged TRF1-T371A (purple bars) and Myc-tagged TRF1-T371D (light yellow bars) from (c). Quantification was performed as described in (b). All P values were calculated using a Student’s two-tailed t test. Standard deviations from three independent experiments are indicated.