Fig. 5

Evaluation of microglial physiological functions in PU-hiMGLs. A Phagocytosis analysis using 1.0-μm Latex beads and fibrillar Aβ1–42, both conjugated with APC fluorescence. Representative images of hiMGLs with and without phagocytosis inhibitor, CCD. Scale bar, 200 μm. Quantification of hiMGLs showing phagocytosis of fluorescence. Two-way ANOVA showed a significant difference between the groups with and without CCD. F (161, 648) = 4.2, p = 0.000835 for Aβ1–42 and F (161, 547) = 3.6, p = 0.000927 for Latex beads, followed by post hoc test ***p < 0.001, n = 3 independent experiments using n = 3 independent clones. B Levels of cytokine gene expression in iMGLs were quantified by qRT-PCR following 3-h LPS treatment. *p < 0.05; **p < 0.01; ***p < 0.001. All data are expressed as mean ± SEM (n = 3 independent clones with n = 2 independent experiments). C The expression level of two key microglial transcription factors (SPI1, IRF8) was evaluated by qRT-PCR after 3-h LPS treatment. ns, not significant; *** p < 0.001. All data are expressed as mean ± SEM (n = 3 independent clones with n = 2 independent experiments). D Composition of the inflammasome (NLRP3, ASC, IL-1β) was confirmed by immunostaining after LPS and Aβ1–42 co-treatment. Scale bar, 100 μm. SEM, standard error of the mean