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. 2015 Aug 5;9:301. doi: 10.3389/fncel.2015.00301

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Copyright © 2015 Smolders, Smolders, Swinnen, Gärtner, Rigo, Legendre and Brône.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Flow cytometry reveals that embryonic microglial cells show a poor expression of activation markers Mac-2, IL1β and iNOS. (A) Gating strategies for the microglial cells. In the whole embryonic cortex cell suspension, a gate was created on the non-debris population (left). Inside this population, single cells were selected (middle) and within this population, the microglial cells were gated based on CX3CR1-eGFP intensity (right). SSC, Side scatter; FSC, Forward scatter. (B) Gating strategies for positive Mac-2, iNOS and IL1β populations. Microglial cell count of representative samples is shown for Mac-2 (left), IL1β (middle) and iNOS (right; full lines) for embryos derived from saline, single poly (I:C) and double poly (I:C) injected mothers. Gates for positive populations were drawn based on the isotype fluorescence intensity (dotted lines). FI, fluorescence intensity. (C) Left panels: at E17.5 only a small percentage of microglial cells shows reactivity for Mac-2. There is no significant effect of poly (I:C) injection on this percentage. Number of embryos tested: Saline N = 5; single poly (I:C) N = 10 and double poly (I:C) N = 10. Middle panels: in control conditions, less than 15% of the microglial cells is positive for IL1β. There is no significant effect of poly (I:C) injection on this proportion. Number of embryos tested: Saline N = 10; single poly (I:C) N = 8 and double poly (I:C) N = 6. Right panels: at E17.5 less than 10% of the microglial cells is positive for iNOS. Poly (I:C) challenge has no significant effect on this percentage. Number of embryos tested: saline N = 5; single poly (I:C) N = 10 and double poly (I:C) N = 10.