Skip to main content
. 2019 Oct 22;33(12):13966–13981. doi: 10.1096/fj.201901460RR

Figure 6.

Figure 6

BI1029539 effect on glutamate-mediated up-regulation of P-gp and mPGES-1 levels in isolated brain capillaries ex vivo. A) P-gp and hmPGES-1 Western blot. Exposing brain capillaries from hmPGES-1 mice to 100 μM glutamate increased P-gp protein expression levels. The glutamate effect on P-gp was blocked by 1, 10, and 100 nM BI1029539. Glutamate also increased hmPGES-1 protein levels in brain capillaries from hmPGES-1 mice; 10 and 100 nM BI1029539 blocked this effect as well. β-Actin was used as protein loading control. B) Representative images of brain capillaries isolated from hmPGES-1 mice after addition of NBD-CSA to determine P-gp transport activity. Glutamate-treated capillaries show increased luminal NBD-CSA accumulation compared with controls. BI1029539 treatment maintained luminal NBD-CSA fluorescence at control levels. Scale bar, 5 μm. C) Image analysis revealed that specific luminal NBD-CSA fluorescence was increased in glutamate-treated capillaries. BI1029539 at all concentrations tested blocked the glutamate effect. For specific luminal NBD-CSA fluorescence, each data point represents means ± sem for 10 capillaries from 1 preparation of 70 hmPGES-1 mice. Units are arbitrary units (au; scale: 0–255). ***P < 0.001, significantly higher than controls.