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. 2014 Feb 3;111(7):2554–2559. doi: 10.1073/pnas.1312570111

Fig. 5.

Fig. 5.

LKB1-dependent HIF1α expression is regulated by mTORC1 and ROS. (A) Immunoblot for LKB1, pS6, p4EBP, and actin protein levels in whole-cell lysates from control (Cre−) or LKB1-null (Cre+) MEFs. (B) Immunoblot for HIF-1α protein levels in A549/Vec or A549/LKB1 cells cultured with (+) or without (−) 25 nM rapamycin for 24 h before cell lysis. Levels of LKB1, pS6, and actin are shown. (C) Relative HIF-1α mRNA expression in MEFs cells from control (Cre−) or LKB1-deficient (Cre+) MEFs treated with 25 nM rapamycin or vehicle control for 24 h. (D) Immunoblot for Raptor and HIF-1α protein levels in whole-cell lysates from control (Cre−) and LKB1-null (Cre+) MEFs treated with control (Ctl) or Raptor-specific (Rapt) siRNA. (E) Relative mean fluorescence intensity (MFI) of DFC-DA staining in LKB1fl/fl cells with (+) or without (−) Cre expression. Cells were treated with or without 10mM N-acetyl cysteine (NAC) for 1 h before ROS measurements. (F) Representative immunoblot of HIF-1α protein expression for cells treated as in E.