Figure 3. Co-expression of wild-type and mutant myocilin does not alter myocilin levels. A: HEK-293T cells were transiently transfected with 200 ng of cDNAs constructs encoding either mutant myocilins E323K or Q368X. These cells were also co-transfected with 200 ng of a cDNA construct encoding wild-type myocilin-HA and 200 ng of either cDNA encoding mutant myocilin-myc E323K or Q368X. Ninety-six hours after transfection, total recombinant myocilin in cell lysates were analyzed by 10% polyacrylamide SDS-PAGE and western blot using the anti-myocilin polyclonal antibody R14T. The black and white arrows indicate the position of full-length myocilin and truncated Q368X myocilin, respectively. Molecular weight markers are shown on the left. B: densitometric quantitation of myocilin detected in A. Error bars represent the S.E. of triplicate experiments.