EMSA of the Atf4 promoter region using palmitate-treated cell extracts. Prior to nuclear extract isolation, HepG2 cells were incubated for 24 h in 0.7 mM palmitate conjugated 2% BSA. EMSA analysis to monitor binding to the CRE site (CGTCA; −164 to −160) was performed as described in Methods section. The first lane marked ‘−’ contains the probe alone and the second lane marked ‘+’ contains the probe and the nuclear extract. The remaining lanes contain antibodies in addition to the probe and the nuclear extract. Arrow represents a complex of ATF4-the DNA probe.