Skip to main content
. Author manuscript; available in PMC: 2012 Oct 7.
Published in final edited form as: Mol Cell. 2011 Oct 7;44(1):134–146. doi: 10.1016/j.molcel.2011.06.038

Figure 5. p62 promotes formation of the active Rag heterodimer.

Figure 5

(A) p62 stabilizes the Rag heterodimer. HEK293T cells were transfected with the indicated cDNAs in expression vectors, cell lysates were prepared, and lysates and Flag-tagged immunoprecipitates were analyzed by immunoblotting to detect the specified ectopic proteins. The graph shows a quantification of the RagB/C dimer formation. Results are represented as means ± standard deviations of three different experiments. *p<0.05. (B) p62 immunoprecipitates from 293 cells, infected with lentiviral shRNA for RagC or RagD, or non-targeting (NT) shRNA, were analyzed by immunoblotting to determine levels of the specified proteins. (C) Cell lysates and RagC immunoprecipitates from WT and p62 KO immortal MEFs reconstituted with p62FL or p62CT were analyzed by immunoblotting to determine levels of specified proteins. (D) Effects of p62 on GTP loading of RagB. The percentage of GTP bound to RagB is indicated for each sample. A portion of cell lysate was taken before immunoprecipitation to probe for expression. (E) p62fl/fl MEFs, infected with GFP or CRE adenovirus, were transfected with Flag-RagBGTP and RagCGDP. Cells were starved for 4 h and restimulated with amino acids for 10 min. Cell lysates were analyzed by immunoblotting to determine levels of the specified proteins. These results are representative of three experiments.