Figure S5.

Effects of H₂R antagonist on the interaction between H₂R and Axin2 or on the WMI in neonatal HI mice. (a) Representative coimmunoprecipitation results showing the effect of histamine, H₂R antagonist cimetidine (Cim), or H₂R agonist amthamine (Amth) on the interaction of H₂R with Axin2 following OGD/reperfusion. (b1 and b2) Neonatal mice were subjected to unilateral common carotid artery ligation plus inhalational hypoxia and received cimetidine administration. Immunohistochemical visualization (b1) and quantification of Iba-1 expression (b2) indicating microglia activation at 14 d after HI. n = 4 mice for each group from two independent experiments. (c1–c3) The H₂R antagonist cimetidine alone or together with H₂R agonist amthamine was administered by a delayed (7–35 d, a–d) approach after HI surgery. Immunohistochemical visualization (c1) and quantification of MBP expression at 35 d (c2) and O4⁺ multipolar differentiating OLs at 21 d (c3). n = 7 mice for each group from three independent experiments. (d1–d3) The H₂R antagonist cimetidine alone or together with H₂R agonist amthamine was administered by a shortened and delayed (7–28 d) approach after HI surgery. Immunohistochemical visualization (d1) and quantification of MBP expression at 28 d (d2) and O4⁺ multipolar differentiating OLs at 14 d (d3). n = 6 mice for each group from three independent experiments. Scale bar, 100 µm. *, P < 0.05; **, P < 0.01; ***, P < 0.001. IB, immunoblot; IF, immunofluorescence staining; IP, immunoprecipitation.