Figure 1.

Schematic of the MYOC eGLuc2 reporter fusion construct. (A) Myocilin is composed of 504 amino acids with a coiled coil myosin (blue/green oval) and OLF-like domains (yellow rectangle) located at the amino and carboxy terminus, respectively. eGLuc2 (pink hexagon) was appended to the C-terminus of MYOC to enable luminescence-based detection. The predicted molecular weight of the construct is 76.5 kDa. Mutants of MYOC used in this study are indicated and their position within MYOC is designated with a red arrow. The vast majority of known MYOC missense mutations occur within the OLF domain. (B, C) HEK-293T cells were transfected with the indicated constructs, and 48 hours after transfection, the conditioned media (B) or soluble cell lysates (C) were analyzed by Western blotting.