The recombinant plasmids pLJM1-CYP1A2, pLJM1-CYP2A6, and
pLJM1-CYP2A13 were identified by PCR and sequencing. The results
showed that pLJM1-CYPs were successfully constructed with no
mutations. CYP2A6, 2A13, and 1A2 expression in transfected cells
was determined by immunoblotting, and the results showed that
a single protein band of each protein was detected in the cells,
whereas the cells transfected with vector alone and blank BEAS-
2B cells showed no bands, suggesting that CYP1A2, CYP2A6, and
CYP2A13 were successfully expressed in BEAS-2B cells (Fig. 1A).
Based on the similar expression level of GFP, several monoclonal
transfected cells were selected and expanded, two of which were
identified by immunoblotting. The results showed that both CYP
proteins and GFP were successfully expressed in all of the monoclonal
cells, in which GFP protein maintained similar expression
levels, including the cells transfected with vector alone (Fig. 1B).
These data further suggested that BEAS-2B cells stably expressing
CYP1A2, 2A6, or CYP2A13 were successfully established