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. Author manuscript; available in PMC: 2015 Jan 1.
Published in final edited form as: Biochim Biophys Acta. 2013 Sep 19;1841(1):10.1016/j.bbalip.2013.09.004. doi: 10.1016/j.bbalip.2013.09.004

Figure 5.

Figure 5

Induction of autophagy is required for tBHQ-induced Nrf2 activation. (A) AML-12 cells were co-transfected with mRFP-GFP-LC3 plasmid and Nrf2 siRNA or scrambled siRNA. The effect of tBHQ on autophagy induction was examined by fluorescence microscopy for puncta formation and Western blot for LC3B-II conversion. All values are denoted as means ± SD from three or more independent batches of cells. Bars with different characters differ significantly, p < 0.05. (B & C) AML-12 cells were treated with tBHQ (50 μM) for 12 h. Autophagy inhibitor, CQ (20 μM) or Bafilomycin A1 (Baf, 100 nM) was added 1 h prior to tBHQ treatment. Nuclear protein and total cellular lysates were subjected to immunoblotting assay for nuclear Nrf2 and keap1, respectively. All values are denoted as means ± SD from three or more independent batches of cells. Bars with different characters differ significantly, p < 0.05.