Figure 7. PSMB5 levels in inducible cells.

A. PSMB5 immunostaining (in red) in inducible, stable RGC5 cells. The cells were induced by Dox for 24 h to express moderate levels of myocilin_WT-GFP (MYOC), myocilin_P370L-GFP (P370L) or myocilin_Q368X-GFP (Q368X). The fusion protein-expressing, green fluorescent cells are shown in insets. There was no green fluorescence, as expected, in non-induced control cells. Note a reduced PSMB5 staining intensity in myocilin-GFP-expressing green cells compared with non-induced cells. Scale bar, 10 µm. B. Western blotting for PSMB5 protein level. RGC5 cells were induced for 24 h to express myocilin_WT (Myoc_WT)-, myocilin_P370L (Myoc_P370L)-, and myocilin_Q368X (Myoc_Q368X)-GFP. Total cell lysate was subject to SDS-PAGE and immunoblotting using anti-PSMB5 or anti-GAPDH. The PSMB5/GAPDH ratios in induced cells relative those of non-induced controls are presented. *, P<0.002 compared to non-induced controls. All experiments were repeated at least 3 times, yielding similar results.