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. 2021 Jun 3;11:11725. doi: 10.1038/s41598-021-91168-5

Figure 1.

Figure 1

CIC expression along the neurogenic lineage in the adult dentate gyrus. (A) A schematic of adult hippocampal neurogenesis. The process of adult hippocampal neurogenesis encompasses four phases with six developmental milestones identifiable through cell morphology and expression of specific protein markers. Radial glia-like (RGL) cells are mostly quiescent stem cells that reside in the subgranular zone. Upon activation, RGL cells give rise to rapidly amplifying type 2a and then type 2b intermediate progenitor cells (IPCs). IPCs differentiate to neuroblasts which eventually exit the cell cycle to become immature and then mature granule neurons. SOX2 and GFAP are expressed by RGL and type 2a cells. DCX starts to be expressed by type 2b cells but is turned off when neurons become mature. (B) A representative image showing CIC, SOX2 and DCX expression in the dentate gyrus of 8-week-old wildtype mice. Scale bar = 50 µm. (C) CIC, SOX2 and DCX expression in the neurogenic subgranular zone. Scale bar = 25 µm. (D) During adult hippocampal neurogenesis, SOX2+ neural progenitor cells express moderate levels of CIC. (E) Minimal CIC expression is observed in DCX+ cells. (F) Mature neurons have the highest levels of CIC. Scale bar = 10 µm. (G) Quantification of relative fluorescence intensity of CIC immunostaining in different cell stages. N = 60 cells from a total of three 8-week-old wildtype mice. (H) CIC is expressed in SOX2+ NES+ RGL cells. Arrow points to a RGL cell. Scale bar = 10 µm. Quantification is shown to the right. N = 59 RGL cells and 80 mature neurons from a total of four 12-week-old control mice. (I) CIC is expressed in SOX2+ GFAP- IPCs. Arrowhead points to an IPC. Scale bar = 10 µm. Quantification is shown to the right. N = 15 IPCs and 51 mature neurons from a total of three 12-week-old control mice. Data are presented in box-and-whisker plots, where centre lines represent medians, box limits represent interquartile ranges, and whiskers represent minimum to maximum data ranges. Statistical analyses were performed with one-way ANOVA with Tukey’s post hoc test (in G) or two-tailed Student’s t-test (H, I). ****P < 0.0001.