Fig. 5.

H₂O₂ activated ectodomain shedding of RAGE. A: NRO cells were incubated with or without H₂O₂ for 1 hr, cells were harvested, and Western blot was analyzed with C-20 anti-RAGE antibody. Lane 1, control without H₂O₂ treatment; lane 2, 100 μM H₂O₂; lane 3, 200 μM H₂O₂. Western blot showing RAGE processed ectodomain shedding by H₂O₂. B: Condition medium was collected, and proteins were precipitated by the TCA method. Western blot was analyzed with N-16 anti-RAGE antibody, and results showed that 45-kDa soluble RAGE was occurred in H₂O₂-treated condition medium. Lane 1, control without H₂O₂ treatment; lane 2, 100 μM H₂O₂; lane 3, 200 μM H₂O₂.