Fig. 4

Knockout of IDO1 ameliorates IBA1+ cells-mediated engulfment of synapse in chronic migraine mice. (A) Representative immunofluorescence images showing colocalization of IDO1 and IBA1 in the ACC of saline- or NTG-treated mice. Scale bars, 20 μm. (B) Quantification of colocalization between IDO1+ and IBA1+ cell numbers in the ACC from saline- and NTG-treated mice (n = 15–16 slices from 4 mice). (C) Representative images of IBA1 immunostaining in the ACC of saline- or NTG-treated mice. Scale bars, 50 μm. (D) Quantification of IBA1+ cells in the ACC of WT and KO mice 2 h after saline or NTG administration (n = 11–14 slices from 4 mice). (E) Representative 3D reconstruction images and Sholl analysis of microglia in the ACC of WT and KO mice with or without NTG-treated. Scale bars, 10 μm (zoom) and 5 μm (rendering). (F) Imaris-based quantification of cell morphometry and total process length of IBA1+ microglia in the ACC from saline- and NTG-treated mice (n = 16–18 slices from 4 mice). (G) Sholl analysis of microglial morphology in saline- or NTG-treated mice (n = 16–18 slices from 4 mice). (H) Representative images and 3D surface rendering of IBA1+ microglia (gray) containing Syn1+ and PSD95+ (red) in the ACC from saline- and NTG-treated mice. Scale bars, 2 μm. (I) Quantification of Syn1+ punta in microglia in mice as indicated in (H) (n = 18–20 cells from 4 mice). (J) Quantification of PSD95+ punta in microglia in mice as indicated in (H) (n = 18–20 cells from 4 mice). The data are shown as mean ± SEM, *p < 0.05, ***p < 0.001, ****p < 0.0001; Student’s t test in (B), one-way ANOVA in (D, F, I, J), two-way ANOVA in (G)