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. Author manuscript; available in PMC: 2014 Feb 8.

Published in final edited form as: Oncogene. 2012 Sep 10;32(32):3765–3781. doi: 10.1038/onc.2012.388

Down-regulation of Nrf2 in human lung A549 cells decreases expression of cytoprotective genes

A) A549 cells were seeded and grown in DMEM that contained 10% FBS for about 24 h before transfer to DMEM containing 0.1% FBS for 16 h. The cells were then treated for 8 h with various doses of either LY294002 or MK-2206 in DMEM containing 0.1% FBS, as indicated, before lysates were prepared and the levels of individual proteins measured by Western blotting. The antibody that recognised both phospho-GSK-3α (Ser-21) and phospho-GSK-3β (Ser-9) was from Cell Signalling (#9331).

B) A549 cells were grown as described above. After serum depletion for 16 h, they were treated for 2 h with various doses of LY294002 or MK-2206 and transferred to fresh DMEM containing 0.1% FBS for a further 6 h before being harvested. Messenger RNA levels were measured by TaqMan RT-PCR.

Down-regulation of Nrf2 in human lung A549 cells decreases expression of cytoprotective genes

A) A549 cells were seeded and grown in DMEM that contained 10% FBS for about 24 h before transfer to DMEM containing 0.1% FBS for 16 h. The cells were then treated for 8 h with various doses of either LY294002 or MK-2206 in DMEM containing 0.1% FBS, as indicated, before lysates were prepared and the levels of individual proteins measured by Western blotting. The antibody that recognised both phospho-GSK-3α (Ser-21) and phospho-GSK-3β (Ser-9) was from Cell Signalling (#9331).

B) A549 cells were grown as described above. After serum depletion for 16 h, they were treated for 2 h with various doses of LY294002 or MK-2206 and transferred to fresh DMEM containing 0.1% FBS for a further 6 h before being harvested. Messenger RNA levels were measured by TaqMan RT-PCR.