Insr deletion in NSCs in vitro decreases NSCs and alters lineage
(A) Induction paradigm of experiment. Cells from ΔNSC-IR WT and ΔNSC-IRKO mice were treated with 0.5 μM 4-OH tamoxifen or vehicle for 24 h.
(B and B′) Neurospheres from vehicle- and tamoxifen-treated cells 5 days after induction.
(C) ΔNSC-IR KO mice have fewer NSCs and a greater proportion of intermediate progenitors within the neurospheres than WT mice (red numbers indicate decreases in fold change, green numbers indicate increases in fold change) (n = 2 independent experiments).
(D and D′) Representative images of tertiary spheres after plating to measure sphere-forming ability.
(E) Average sizes of tertiary tdTomato red+ neurospheres.
(F) Percentage of tertiary tdTomato red+ neurospheres per field were reduced in ΔNSC-IRKO mice (unpaired t test, p < 0.0001; n = 3 independent experiments).
BNAP, bipotential neuron-astrocyte progenitor; GRP, glial restricted progenitor; MP, multipotent progenitor; NSC, neural stem cell; PFMP, platelet-derived growth factor-fibroblast growth factor 2 (PDGF-FGF2)-responsive MP.