FIG. 6.

MondoA can regulate glycolytic genes directly. (A) The activities of the wild-type (WT) and double CACGTG E-box mutant (mE1 + mE2) LDH-A luciferase reporters were measured in C2C12 cells expressing the indicated combinations of ΔN237MondoA (MondoA) and NLS-Mlx (Mlx) or the corresponding empty expression vectors. (B) Schematic diagrams of the LDH-A, HKII, and PFKFB3 promoters. Exons (E) and transcriptional start sites (+1) and locations of CACGTG E-box elements are indicated. Lines above the promoters indicate the regions amplified by PCR. (C) Real-time PCR assay measuring enrichment of ChIPs from C2C12 cells expressing ΔN237NLSMondoA performed with MondoA, c-Myc, or Gal4p antibodies. Immunoprecipitated DNA was analyzed by real-time PCR with primers specific to the E-box-containing regions of the mouse LDH-A, HKII, and PFKFB3 promoters. Measurements are expressed as fold enrichment over the amount of an upstream region of the PFKFB3 promoter that does not contain a CACGTG site. Each value is the average (±the standard error) from two independent biological replicates.