Fig. 7.

AAR-induced cJUN binding to the ATF3 promoter is independent of ATF4. HepG2 cells stably expressing a doxycycline (DOX)-inducible shRNA against either cJUN (A) or ATF4 (B) were treated with DMEM or DMEM containing 2 mM HisOH for 6 h after the shRNA expression was induced by DOX for 48 h. Nuclear protein extracts were assayed for transcription factor binding by DAPA with a DNA oligonucleotide (nt −104/+27) containing both the ATF/CRE and CARE sites of ATF3 promoter region. An aliquot of the nuclear extract was subjected to immunoblotting with the indicated primary antibody (“Input”). Equal loading of the samples was established by Fast Green staining of the blot (not shown). The blots shown are representative of multiple experiments.