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. 2014 Apr 14;9(4):e92845. doi: 10.1371/journal.pone.0092845

Figure 1. Turnover of the endogenous myocilin protein and β-catenin.

Figure 1

A. Human TM cells from different donors were treated with CHX (5 µg/ml) and monesin (2 µM). Cells were harvested at various time points thereafter and the level of endogenous myocilin was determined by Western blotting. The GAPDH level was used for protein loading control. The level of the endogenous myocilin was dropped to about 50% 3 h after CHX and monesin treatment, indicating that myocilin is a short-lived protein. One representative experiment is presented. Ratios between levels of the endogenous myocilin and those of GAPDH are presented (left panel), which were plotted against time to estimate the myocilin half-life (right panel). B. The turnover of the endogenous myocilin in RGC5 cells. Experiments were done as described in A. The half-life was estimated to be about 3 h. C. The turnover of β-catenin, a known short-lived protein, in RGC5 cells. Its half-life was found to be approximately 6 h, comparable to that reported previously [41]. Experiments were repeated at least 3 times, yielding similar results.