FIGURE 5.

Expression of CEL14R and CEL MODY in AR42J cells transduced with adenovirus constructs. A, total RNA was isolated from cells transduced with a control or a CEL14R- or CEL MODY-containing adenovirus. The amount of mRNA encoding human CEL and GAPDH was determined by qRT-PCR as described under “Experimental Procedures.” There is no significant difference in the amount of mRNA encoding CEL14R and CEL MODY (n = 6). B, the cells were transfected with the indicated adenovirus as described under “Experimental Procedures.” Samples were obtained from the medium and from the intact cells. In separate experiments, the intracellular CEL was separated into soluble and insoluble fractions as described under “Experimental Procedures.” The proteins in each sample were separated by SDS-PAGE, and immunoblotting was performed with CEL antiserum. C, the amount of immunoreactive protein was quantified by densitometry and presented in bar graphs. The medium and soluble fractions contained significantly more CEL14R (p = 0.001). The total intracellular sample and the insoluble fraction contained significantly more CEL MODY (p = 0.001) (n = 8). D, cells were transduced with the indicated adenoviral constructs, and the insoluble fraction was isolated. The samples were separated by SDS-PAGE in the presence or absence of β-mercaptoethanol, and CEL was detected by immunoblotting. Representative gels from three separate experiments are shown. Error bars represent S.D.