Fig. 4

P2Y12R affected NTG-induced microglial morphological alterations and iNOS production in the TNC. a Iba-1 immunostaining in the TNC following NTG, MRS2395, or clopidogrel administration. Scale bar: 20 μm. b Magnified images of Iba-1 in the area enclosed in the white dotted line frame in a. c, f Quantification of Iba-1-positive cells (c) and Iba-1 immunoreactivity (f) in the TNC. Data are presented as mean ± SEM; n = 4 per group; six sections from each mouse; *p < 0.05, **p < 0.01 compared with the sham group. d, e Quantitative analysis of the total length of initial processes (d) and the mean length of initial processes per microglia (e). Data are presented as mean ± SEM. A total of 340 cells (71 cells in the sham group, 68 cells in the NTG group, 62 cells in the NTG + DMSO group, 67 cells in the NTG + MRS2395 group, 72 cells in the NTG + Clop15 (15 mg/kg) group) were analyzed. *p < 0.05, **p < 0.01, and ***p < 0.001 compared with the sham group; ^#p < 0.05 and ^##p < 0.01 compared with the NTG group. g Representative immunoblots of iNOS expression in the TNC at various time points after NTG administration. The band intensities are presented relative to those of β-actin. Quantification of iNOS expression indicated a significant increase on day 9 after NTG injection. Data are presented as mean ± SEM; n = 6 per group; *p < 0.05 compared with the sham group. h Western blot showing that the P2Y12R inhibitors MRS2395 and clopidogrel (15 mg/kg) reduced the production of iNOS induced by NTG in the TNC. Values are presented as mean ± SEM; n = 6 per group; **p < 0.01 compared with the sham group; ^###p < 0.001 compared with the NTG group