Activation of the Nrf2 Pathway by Inorganic Arsenic in Human Hepatocytes and the Role of Transcriptional Repressor Bach1
Figure 5
NaAsO2-induced nuclear import of Nrf2 is accompanied with the nuclear export of Bach1. (a) Chang human hepatocytes were treated with 25 μmol/L of NaAsO2 for different time interval (0.5, 1, 2, and 4 h). Nuclear and cytosolic proteins were extracted separately to perform the western blot assay. Representative immunostained bands of nuclear (left) and cytosolic (right) proteins illustrated the changes of subcellular localization of Nrf2 and Bach1. (b) and (c) were quantitative analysis of nuclear and cytosolic Nrf2 and Bach1 proteins as shown in (a). β-actin and lamin B were used as internal control, accordingly. (e) The subcellular location of Bach1 was detected by immunofluorescence. Chang human hepatocytes were treated with 25 μmol/L of NaAsO2 for 4 h, immunostained with anti-Bach1 and TRITC-conjugated second antibodies (red, far left) and counterstained with DAPI to show the nucleus (blue, middle). The far right panels represented the overlay of Bach1 and DAPI fluorescence images. The results shown here were representative of three separate experiments.
