Skip to main content
. 2014 Apr 16;9(4):e95393. doi: 10.1371/journal.pone.0095393

Figure 3. UA does not inhibit the associations between mTOR and Raptor or Deptor.

Figure 3

(A), Deptor-suppressed and scrambled C2C12 cells were serum starved and pretreated with or without 50 µM UA for 60 min, followed with or without 10 mM leucine (Leu) for 60 min. The phosphorylation of S6K, 4EBP1 and the protein levels of S6K, 4EBP1, Deptor, and Tubulin were determined by Western blot using specific antibodies. (B), S6K phosphorylation in (A) was semi-quantified using the NIH Image J Program and normalized with S6K protein levels. (C), C2C12 cells were serum starved and pretreated with or without leucine (Leu) for 60 min, followed with or without 50 µM UA for 60 min. mTOR was immunoprecipitated from cell lysates using an anti-mTOR antibody and the co-immunoprecipitaed Raptor or Deptor was determined by Western blot. Differences between groups were examined for statistical significance using ANOVA. Data are presented as mean ± S.E.M. from three independent experiments. NIg, normal immunoglobulin. *, P<0.05, **, P<0.01, and ***, p<0.001. N, no addition.