FIG. 4.

Changes in nuclear ChREBP in response to verapamil and CyA. INS-1 cells were incubated in the presence or absence of verapamil (50 μmol/L) for 24 h, and ChREBP levels were analyzed by Western blotting in whole cell extracts (A) or nuclear fractions (B) as detailed in Research Design and Methods. C: INS-1 cells were transfected with the FL-TXNIP promoter reporter plasmid (0.4 μg/well) together with pRL-TK control plasmid (20 ng/well) using DharmaFECT Duo transfection reagent (1 μL/well) and incubated in the presence or absence of the calcineurin inhibitor CyA (5 μmol/L) for 24 h. Bars represent luciferase activity (mean ± SEM) of three independent experiments. D: INS-1 cells were incubated in the presence or absence of CyA (5 μmol/L) for 24 h, and ChREBP levels were assessed in the nuclear fractions (n = 3).