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Ngfr regulates neurogenic response in DG astrocytes through suppression of Lcn2/Slc22a17 activity. a Single cell transcriptomics strategy for isolating Ngfr-mCherry positive astrocytes and comparing the transcriptomics profiles to non-transduced astrocytes or control astrocytes. The comparison showed differential expression of 10 genes in common as shown in the heat map. b Expression of selected genes on tSNE plots. c Violin plots for selected genes. d Immunostaining for Lcn2 and mCherry with DAPI counterstain in control and Ngfr+ mouse hippocampal astrocytes. e Quantification of Lcn2 signal intensity normalized to cell numbers. f Immunostaining for Lcn2, Gfap and mCherry in LV16 Ngfr-transduced DG. Lcn2-positive and mCherry positive astrocytes do not overlap. g tSNE plots for two receptors of Lcn2: Lrp2, Slc22a17. h Gfap and Lcn2 immunostaining indicates increased Lcn2 and gliosis in APP/PS1dE9 mice. i Heatmap showing the changes in astrocyte A1 state markers upon Ngfr+ transduction. Astrocyte cluster 6 clusters showing expression of C1qa, Gfap, C4b, Il1b, and Serpina3n in control and Ngfr+ states. j Injection, BrdU treatment and analyses scheme of the effect of Lcn2 on proliferating GFAP cells, and immunohistochemical staining for BrdU, GFAP and DAPI. k Quantification graph for BrdU-GFAP double positive cells. l Schematic representation of the cell-type specific functional knockdown of Slc22a17 in astrocytes. m mCherry, Dcx, BrdU immunostaining with DAPI counterstain in SGZs transplanted with Lv13+control morpholino, Lv13+Slc22a17 morpholino and Lv16+control morpholino-treated astrocytes. Lower panels: DAPI omitted from upper panels n Quantification of Dcx, mCherry and BrdU triple positive cells in i. Slc22a17 knockdown mimics Ngfr transduction. n = 3. Scale bars equal 25 μm. Error bars represent standard error of the means.
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