Figure 4.

C-DIM/NR4A1 ligands and metformin induce glucose uptake and gene products associated with this response. C2C12 cells were treated with DMSO and (A) DIM-C-pPhOH, (B) DIM-C-pPhOH-3,5-Br₂, (C) DIM-C-pPhOH-3-Cl, and (D) DIM-C-pPhOH-3-Cl-5-OCH₃ for 24 hours, and whole-cell lysates were analyzed by western blots. C2C12 cells were treated with (E) DIM-C-pPhOH-3-Cl-5-OCH₃ and (F) metformin for 6, 12, and 24 hours, and whole cell lysates were analyzed by western blots. (G) C2C12 cells were treated with C-DIM compounds and metformin for 24 hours, and glucose uptake in the cells was determined using the Glucose Uptake-Glo Assay Kit (Promega). Results are expressed as means ± SE for at least three replicated determinations, significant (P < 0.05) glucose uptake is indicated (*), and glucose uptake in control cells was 1.0. (H) DIM-C-pPhOH-3-Cl-5-OCH₃ and metformin treatment also induced GLUT1 expression as determined by western blot. (I) C2C12 cells were treated with DIM-C-pPhOH-3-Cl-5-OCH₃ and metformin alone or in combination with 5 mM glutathione (GSH) for 12 hours, and whole-cell lysates were analyzed by western blots for sestrin 2. Met, metformin.