Figure 6.
Responses of microglia to ZIKV infection in brain organoids
(A) Representative phase-contrast images showing the morphology of day 75 microglia-containing organoid after 3 and 7 days of ZIKV or mock infection. Scale bars, 200 μm.
(B) Quantitation of infectious ZIKV particles in the culture medium by plaque assay. n = 3, three independent times infections, each time, 8–12 organoids from two hiPSC lines. Data are presented as mean ± SEM.
(C) Representative images showing E protein expression in day 75 organoids after 3 days of ZIKV or mock infection. Scale bars, 200 μm.
(D) Representatives of IBA1+/CD68+ microglia in day 75 organoids after 3 days of ZIKV or mock infection. Scale bars, 10 μm.
(E) Quantification of the number of endpoints and process length of IBA1+ microglia in day 75 organoids after 3 days of ZIKV or mock infection (n = 3, 3 independent times infections, each time, 8–12 organoids from two hiPSC lines). Student's t test. ∗p < 0.05. Data are presented as mean ± SEM.
(F–H) (F) qPCR analysis of IL-6, IL-β, and TNF-α mRNA, (G) IFNAR1 and IFNAR2 mRNA, and (H) CX3CR1, C3, and CR3 mRNA expression in day 75 organoid after 3 days of ZIKV or mock infection. n = 3, three independent times infections, each time, 10 organoids from two hiPSC lines. Student's t test. ∗p < 0.05, ∗∗p < 0.01. Data are presented as mean ± SEM.
(I and J) (I) Representative 3D reconstruction and (J) quantification of PSD95+ puncta engulfment in CD68+ microglial phagolysosomes in day 75 organoids after 3 days of ZIKV or mock infection. Scale bars, 5 or 2 μm as indicated. n = 3, three independent times infections, each time, 8–12 organoids from two hiPSC lines. Data normalized to engulfment in the mock control. Student's t test. ∗∗p < 0.01. Data are presented as mean ± SEM.