Fig. 9: In vivo and in vitro functional validation of the downregulated PTEN pathway and upregulated mTOR pathway as key pathway mediating the increased neurogenesis in the MG-Alk5 iKO mice. | Nature Communications

Fig. 9: In vivo and in vitro functional validation of the downregulated PTEN pathway and upregulated mTOR pathway as key pathway mediating the increased neurogenesis in the MG-Alk5 iKO mice.

From: Inhibition of TGF-β signaling in microglia stimulates hippocampal adult neurogenesis and reduces anxiety-like behavior in adult mice

Fig. 9: In vivo and in vitro functional validation of the downregulated PTEN pathway and upregulated mTOR pathway as key pathway mediating the increased neurogenesis in the MG-Alk5 iKO mice.

A, B In vivo animal models used and experimental timeline. C Representative images of Cx3cr1CreER-ALK5WT/WT or Cx3cr1CreER-ALK5fl/fl treated with either Vehicle (VEH) or Rapamycin (RAPA) showing immunohistochemistry staining for DCX and IBA1 3 weeks after Tamoxifen treatment. Yellow Arrowhead shows DCX+ cells migrated out of the SGZ inner layer. D Percentage of total DCX+ cells compared to the wildtype mean and E percentage area of dendritic arborization compared to the wildtype mean. F Percentage of migrated DCX+ cells relative to the total number of DCX+ cells per mouse. G, H Representative images of WT or iKO mice treated with either VEH or RAPA showing immunohistochemistry staining for DCX and pS6 3 weeks after TAM treatment. D–G WT VEH n = 7, KO VEH n = 4, WT RAPA n = 5, and KO RAPA n = 4 (ns=not significant (D–G); *** p < 0.001, ** p = 0.01 D; ** p = 0.003, *** p < 0.001 E; * p = 0.042, *** p < 0.001 F; * p = 0.048, ** p = 0.01, *** p < 0.001 G). The sex of each animal is represented by open circles (females) and open triangles (males). I In vitro experimental model and timeline for primary adult NSCs and microglia (P2ry12CreER/CreER-tdTomato) 3D co-culture. J Representative images of a differentiated neuroimmune 3D coculture sphere showing microglial tdTomato, GFAP and TUJI immunostaining (maximum projection from Z stacked confocal images). K,L Representative images of the 3D coculture spheres quantification of the TUJI immunoreactive density in the spheres after differentiation with indicated microglia genotypes and VEH or RAPA treatment (WT VEH n = 4, iKO VEH n = 6, WT RAPA n = 6, iKO RAPA n = 4) (ns=not significant, * p = 0.0352, *** p = 0.001 K). M A proposed model of mechanistic action through microglia onto adult-born neurons. Mean ± SE. Scale bar = 100 µm, Two-way ANOVA test, two-sided, with Tukey post hoc pairwise comparison was used for statistical analysis for all panels. Panel A, B,I and M were created in BioRender. Luo, A. (2026) https://BioRender.com/sxk2w6x. Source data are provided as a source data file.

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