Figure 4

Cellular localization of CXR in transiently transfected LMH cells. A, Full-length CXR, CXR with GFP attached at its C-terminus or at its N-terminus or N-terminal GFP-CXR fusion protein mutated in its xenochemical response signal (XRS) at positions 356, 359 and 362 were expressed in CV-1 cells together with a reporter plasmid containing the CYP2H1 264-bp PBRU. After transfection, the cells were treated with either vehicle, 400 μM PB or 10 μM clotrimazole before cells were lysed and analysed for reporter gene expression. Values are the average of three independent experiments and error bars represent standard deviations. B–F, LMH cells were transfected with either pEGFP-vector alone (B), expression vector for N-terminal GFP-CXR fusion protein treated with vehicle (C), 400 μM PB (D) or 0.1 μM okadaic acid (E) for 16 hours or GFP-CXR fusion protein with the xenochemical response signal mutation as described in Fig. 4A (F). Cells were stained with 300 nM DAPI in PBS and analysed for DAPI and GFP-specific light emissions at 461 nm and 507 nm using excitation wavelengths of 358 nm and 488 nm, respectively. Size bars stand for 20 μm.