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. 2013 Dec 13;8(12):e83318. doi: 10.1371/journal.pone.0083318

Figure 2. Decreased levels of ubiquitinated proteins in U87MG cells that express PTEN.

Figure 2

The same U87MG cells from Figure 1 were incubated for 3 h in KH without (A) or with (B) amino acids and insulin. Where indicated, the cells were treated with the proteasome inhibitor MG132 (50 µM). Extracts (75 µg protein) of the cells were subjected to SDS-PAGE and analyzed by Western blot using two different antibodies that recognize ubiquitinated proteins (FK1 and FK2) and, as a loading control, an antibody that recognizes actin. The amount of ubiquitinated proteins (ubiq. prot.) was quantified by densitometry and normalized to the levels of actin. Results, which are means ± SD of three independent experiments, are shown below the representative Western blots. Differences from the corresponding values (with or without MG132) in mock-treated cells were found to be significant at **p<0.01 and ***p<0.005, respectively.