Figure 4.

iMG cells exhibit appropriate physiological responses to LPS and IFN-γ challenge and are able to engulf microspheres or fibrillar Aβ

(A) qRT-PCR analysis of inflammation-related gene expression in N2-iMG cells after LPS/IFN-γ stimulation for 6 h. Data are means ± SEM (TNF-α, n = 10; IL-6, n = 14; iNOS, n = 14; IL-10, n = 4; CD68, n = 13). ^∗∗p < 0.01, ^∗∗∗∗p < 0.0001 compared with the mock-treatment group (ratio paired t test).

(B and C) Representative spinning-disc confocal microscopy images of phagocytosis by N2-iMG cells (day 9) of latex beads (B, red) or TAMRA-labeled fibrillar Aβ (fAβ) (C, red). Cells were incubated with substrates for 1 h. The plasma membrane and nuclei of the live cells were stained with CellMask deep red (green) and Hoechst 33,342 (blue), respectively. The z-axis images at the vertical and horizontal yellow lines were extracted from 3D images, and indicate right and bottom positions, respectively. Scale bar, 10 μm.

(D and E) Flow cytometry analysis of fluorescent microsphere bead (D) or TAMRA-fAβ (E) uptake by N2-iMGs at day 9.

(F and G) Quantitative results for percentage of CD11b⁺ cells with fluorescent beads (F) or fAβs (G). Data are means ± SEM (n = 3–6 independent experiments). ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001, ^∗∗∗∗p < 0.0001 by one-way ANOVA using Tukey's multiple comparisons test. Cyto D, cytochalasin D.