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. Author manuscript; available in PMC: 2017 Jul 26.
Published in final edited form as: Nature. 2017 Jan 18;541(7638):481–487. doi: 10.1038/nature21029

Figure 1. Serum-free culture model for A1 reactive astrocytes.

Figure 1

a, Heat map of reactive transcripts. Csf1r−/− mice (which lack microglia) fail to produce A1 astrocytes following LPS injection. LPS-activated microglia, or a combination of Il-1α, TNFα, and C1q are able to induce A1s in culture. b, Cytokine array analysis of LPS-activated microglia conditioned media (MCM) with increases in Il-1α, Il-1β and TNFα (Il-1β was not A1-specific). c, Western blot analysis of LPS-activated MCM for C1q protein. d, TGFβ was able to reset A1 reactive astrocytes to a non-reactive state. e, Individual knock-out (Il-1α−/−, TNFα−/−, C1q−/−), double (Il-1α−/−TNFα−/−), and triple knock-out (Il-1α−/−TNFα−/−C1q−/−) mice fail to produce A1s following LPS injection. Mice treated with Pexidartinib (PLX-3397) for 7 days to deplete 95% of microglia (Extended Data Fig. 1) still respond to LPS by producing A1s. f, Representative phase and fluorescent immunohistochemistry micrographs for GFAP and AQP4 of control and A1 reactive astrocytes. g, Western blot analysis of GFAP protein in cultured astrocytes showing approximate 3-fold increase in A1s compared to control. h, Measurements of cross-sectional area of astrocytes stained with GFAP. n = 6–8 for each experiment. * p < 0.05, one-way ANOVA. Error bars indicate s.e.m. Scale bar: 50 μm.