Inhibition of the ALP stabilizes a UPS substrate.

NRVMs were cultured and infected with Ad-GFPu/Ad-RFP viral mixture as described in Figure 4. Forty-eight hours later, the cells were treated with 3-MA (2 mM, A and B) or BFA (10 nM, C and D), or respective vehicle control (CTL) for 12 hrs; then, the treatment of cycloheximide (CHX, 50 µM) was initiated. Immediately before (0 min) or 5, 10, 15, 30, 60, and 120 min after CHX administration, the cells were harvested for western blot analyses for GFPu and RFP. Representative western blot analysis images (A, C) and densitometry data pooled from 3 biological repeats (B, D) are shown. *p<0.05, **p<0.01 vs. CTL.