Figure 6.

ERRγ regulates the FAO via Cpt1b in chemoresistant cancer cells. (A~B) Relative FA uptake (A) and FA β oxidation rate (B) between HepG2 and HepG2/ADR cells; (C~D) Relative FA uptake (C) and FA β oxidation rate (D) in cells transfected with sh-Con or sh-ERRγ; (E) Cell proliferation measured by CCK-8 kit in HepG2 cells pre-transfected with vector control or pcDNA/ERRγ for 6 h and then treated with or without Dox (1 μM) combined with or without ETO for 24 h; (F) mRNA levels of FAO-related genes measured by qRT-PCR in HepG2 and HepG2/ADR cells; (G) mRNA levels of FAO-related genes measured by qRT-PCR in HepG2/ADR cells transfected with sh-Con or sh-ERRγ; (H) Cell proliferation measured by CCK-8 kit in HepG2/ADR cells pre-transfected with sh-Con or sh-ERRγ and then transfected with vector or a Cpt1b expression construct, followed by further treatment with Dox (10 μM) for 24 h; (I~J) ATP (I) and FA β oxidation rate (J) measured in HepG2/ADR cells transfected with sh-Con or sh-ERRγ with further transfection with vector or a Cpt1b expression construct for 24 h; (K) Nucleotide sequences of ERREs in CPT1B and the mutated (GACCTTG to AGAACCG) nucleotides in pGL3-CPT1B-Mut-Luc vector; (L) ChIP assay measuring ERRγ binding to CPT1B promoter in both HepG2 and HepG2/ADR cells; (M) Dual-luciferase reporter gene assay performed in HepG2 and HepG2/ADR cells transfected with pGL3-CPT1B-WT-Luc or pGL3-CPT1B-Mut-Luc; (N) Dual-luciferase reporter gene assay performed in HepG2 cells transfected with pGL-ABCB1-WT or pGL3-CPT1B-Mut reporter with or without pcDNA/ERRγ for 12 h and then further treated with or without BAY 11-7082 for 12 h; (O) Model for ERRγ-regulated FAO via Cpt1b in chemoresistant cancer cells. Data were presented as means ± SD from three independent experiments. *p< 0.05.