Extended Data Fig. 3: Loss of Shisa9 promotes antiviral immune responses in microglia and astrocytes.
From: Modulation of virus-induced neuroinflammation by the autophagy receptor SHISA9 in mice
(a) Real-time quantitative polymerase chain reaction (qPCR) analysis of the expression of Shisa9 in wild-type (WT) microglia and astrocytes infected with HSV-1 (MOI = 1) for the indicated time periods. (b) Expression scatterplots of cell type markers in population from (Fig. 2a). (c) The expression of Shisa9 in different cell types is analyzed from the scRNA data of day 3 and day 7. (d) The proportion analysis of astrocytes in different samples. (e) The expression of inflammatory genes in 8 subclusters of astrocytes. (f-g) Pseudotime analysis of astrocytes from WT mice compared to Shisa9−/− mice. (f) The expression of Shisa9 in WT astrocytes along with pseudotime axis. (g) Pseudo time-line differential genes. (h) The expression of inflammatory genes was presented as a heatmap with pseudotime axis. For every time point, each group is a mix of brain tissues from 5 mice in Extended Data Fig. 3b–h. (i) qPCR analysis of indicated interferon-stimulated genes (ISGs, Mx1, Oas1, Ifit1 and Ifit2) or chemokines (Ccl2, Ccl5, Ccl7, Cxcl9 and Cxcl10) in microglia or astrocytes transfected with control siRNA or Shisa9 siRNA, followed by HSV-1 (MOI = 1, 48hrs) infection. (j and k) Immunofluorescence (j) and quantitation (k) for IFNβ or IL-6 (red) on Iba1+ microglia or GFAP+ astrocytes (green) to identify the source of indicated cytokines in the brains of WT and Shisa9-/- mice at day 0 (HSV-1 uninfected, UI). Scale bars (j), 100 μm. Data In (a and i), data are presented as mean ± SEM, P-values were determined by unpaired two-tailed Student’s t test of 3 independent biological experiments, ns: not significant. In (k), mean ± SD of 8 mice. P-values were determined by unpaired two-tailed Student’s t test.
