Single cell profiling of CD11b+ cells from mouse cortex following astrocyte overexpression of mCherry-CAAX or Hevin
Astrocyte derived Hevin regulates formation and maintenance of excitatory thalamocortical synapses in the developing mouse cortex. However, the role of astrocyte secreted factors on other cell types is still understudied. To interrogate the response of microglia to the release of astrocye derived Hevin, astrocytes were virally transduced with AAVs to overexpress Hevin or mCherry-CAAX. Overall design: P0-P1 mice were intracortically injected with AAVs to overexpress Heivn or mCherry-CAAX under the control of the GfaABC1D promoter to restrict expression to astrocytes. Microglia were then isolated from cortex of these mice at P15 using Magnetic Activated Cell Sorting (MACS) for CD11b+ cells. These CD11b+ cells were then used as input for single cell library preparation using the PIPseq v4.0PLUS kit from FluentBio. Libraries were then sequenced using Illumina NovaSeq 6000.