Fig. 5

PEG-CeNP treatment modulated microglial polarization via inhibiting ROS-induced NF-κB p65 translocation. a Flow cytometric analysis of microglia isolated from the ipsilateral hemisphere at 3 days post ICH. Representative gating strategy for microglia (CD45^low+CD11b⁺), macrophages (CD45^high+CD11b⁺), M1 microglia (CD16/32⁺CD206⁻,CD45^low+CD11b⁺), and M2 microglia (CD16/32⁻CD206⁺,CD45^low+CD11b⁺). b Quantification of CD16/32⁺CD206⁻ microglia, n = 6/group, *p < 0.01 versus ICH + vehicle. c Quantification of CD16/32⁻CD206⁺ microglia, n = 6/group, * p < 0.05 versus ICH + vehicle. d Mean fluorescence intensity of CD206 in ICH + vehicle (gray) and ICH+ CeNP (green) group. e, f Representative Western blot bands showed the expression of NF-κB p65 and phosphorylated NF-κB p65 (P-p65) in brain tissues at 3 days after ICH, n = 6/group, * p < 0.05 versus Sham, # p < 0.05 versus ICH + vehicle. g ROS levels in cultured microglia, n = 6/group, * p < 0.05 versus Control, # p < 0.05 versus H₂O₂ + vehicle. h Representative Western blot bands showed the expression of IκB-α in cultured microglia, n = 6/group, * p < 0.05 versus Control, # p < 0.05 versus H₂O₂ + vehicle. i Representative images of NF-κB p65 and Iba1 double immunostaining in cultured microglia. Lens: × 400; Scale bar: 25 μm. j, k Representative Western blot bands showed the expression of nuclear and cytoplasm NF-κB p65 in cultured microglia, n = 6/group, * p < 0.05 versus Control, # p < 0.05 versus H₂O₂ + vehicle