Figure 9. Effect of dominant-negative A-USF expression on HO-1 protein expression.

HK-2 cells were transiently transfected with empty vector (upper panel) or the dominant negative, HA-tagged A-USF vector (lower panels). Cells were stimulated 24 h after transfection with haemin (5 μM) for 4 h. Dual-label immunofluorescence was performed using Alexa 594-conjugated anti-HA (Molecular Probes) and anti-HO-1 (Stressgen Biotechnologies) antibodies for detection of A-USF and HO-1 respectively. HO-1 was detected using a goat anti-rabbit FITC-conjugated secondary antibody (Jackson Immunoresearch Laboratories). Results are representative of two independent experiments. DAPI, 4,6-diamidino-2-phenylindole.