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. 2013 Jun 25;41(16):7683–7699. doi: 10.1093/nar/gkt563

Figure 6.

Figure 6.

Effect of leucine concentration on the transcriptional activation of members of the three classes of ATF4-dependent autophagy genes. (A) Immunoblot analysis of ATF4, CHOP protein contents and MAP1LC3B processing, from MEFs incubated for 16 h in medium containing different leucine concentration. The control medium contained 420 µM leucine. The LC3-II/LC3-I ratio was calculated based on densitometry analysis of both bands using Image J software. Induction of expression of (B) Chop and Atf4, (C) p62, Nbr1 and Atg7 where both ATF4 and CHOP were bound to the AARE, (D) Atg10, Gabarap and Atg5 where CHOP was bound to CHOP-RE without interacting with ATF4 and (E) CHOP-independent autophagy genes where ATF4 was bound to AARE without interacting with CHOP, in response to different leucine concentrations. MEFs were incubated for 16 h in media containing 420 or 150 (black), 100 (gray), 70 or 30 µM (white) and harvested for the mRNA content analysis. The graphs show means ± S.E.M. of three independent experiments. t tests have been performed to compare the means. The asteriks indicate a P value of ≤ 0.05 relative to the 420 µM medium value.