Figure 2. Characterization of the signal sequence cleavage of myocilin. A: HTM cells were transduced with Ad-myocilin-FLAG and myocilin in cells (lane 1), and culture media (lane 2) were separated by SDS–PAGE, transferred to the membrane, and loaded into a protein sequencer. All myocilin bands yielded the same results with respect to their NH₂-terminal amino acid sequences. B: HTM cells were transduced with Ad-FLAG-myocilin and myocilin in the cells (lane 2), and culture media (lane 3) was detected by western blot analysis using anti-myocilin antibody (upper panel) or anti-FLAG antibody (lower panel). Myocilin-FLAG (Figure 1A, lane 2) was included as a control (lane 1).