Fig. 9.

Recombinant sRAGE increased p-eNOS expression and decreased F-actin formation in HBMECs exposed to glia-conditioned media or SAH-patient's CSF. (A) Time course of p-eNOS (ser1177) and eNOS protein expression by Western immunoblot in HBMECs exposed to 10 ​% SAH-patient's CSF. The p-eNOS and eNOS protein levels are normalized to GAPDH loading control. (B) Representative immunofluorescence images of F-actin (green, visualized with phalloidin staining) and p-eNOS (red) in HBMECs and stimulated with glia-conditioned media or SAH-patient's CSF for 12 ​h. Arrows indicate stress fibers formed. Bar ​= ​50 ​μm. (C) Immunoblot detection and quantification of (D) p-eNOS (ser1177), (E) eNOS, (F) cleaved-caspase3 and GAPDH in HBMECs treated with sRAGE for 12 ​h. The levels of p-eNOS (Ser 1177) and eNOS were increased and cleaved caspase-3 was attenuated in the presence of sRAGE in HBMECs exposed to glia-conditioned media or SAH-patient's CSF. Expression for each sample was normalized with corresponding GAPDH level. Data are expressed as means ​± ​SD. ∗P ​< ​0.05, ∗∗P ​< ​0.01, ∗∗∗P ​< ​0.001 (n ​= ​3 in each group).