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. 2014 May 1;289(24):16790–16801. doi: 10.1074/jbc.M114.549717

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© 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 6. — A reduction in the level of total or surface megalin decreases the PKB pathway and overactivates the ERK/mTORC1/S6K pathway. A, the luminal expression levels of megalin were assessed by surface biotinylation (n = 3). B–E, LLC-PK1 cells stably transfected with shRNA for megalin were grown on six-well plates and kept overnight in serum-depleted medium. B, total and luminal expression levels of megalin (n = 6). C, PKB, mTOR, S6K, ERK, and RICTOR phosphorylation (n = 6). D, densitometry analysis. PKB phosphorylation at Ser-473 (first bar); mTOR phosphorylation at Ser-2448 (second bar) and Ser-2481 (third bar); S6K phosphorylation at Ser-371 (fourth bar), Thr-389 (fifth bar), Thr-421/Ser-424 (sixth bar); ERK phosphorylation at Thr-202/Tyr-204 (seventh bar) and RICTOR phosphorylation at Thr-1135 (eighth bar). E, mTORC1 and S6K activities (n = 6). The results are shown as means ± S.E. *, p < 0.05 versus control.