Figure 2. Targeting CPT1 or Fatty Acid Mobilization Diminished the NSC Pool.

(A and B) Silencing Cpt1a diminished the NSC pool. Mouse embryos were electroporated at E12.5 and sacrificed at E15.5. In the rescue group (Cpt1a shRNA + CPT1A), a plasmid for expressing shRNA-resistant mouse Cpt1a cDNA was co-electroporated with Cpt1a shRNA and EGFP plasmid. (A) Representative confocal images. Areas of the ventricular zone (Pax6⁺ layer) are shown at higher magnification in bottom panels. Scale bars: 50μm. (B) Quantifications. (C–F) Etomoxir induces increased differentiation of NSCs. (C) Neocortical cells used for FAO assay. Dissociated neocortical cells of E11.5 mouse embryos were cultured overnight, and then used in [¹⁴C]-oleate oxidation assay for assessing FAO activity. Most cultured cells express the NSC marker Nestin. DAPI labels the nucleus of all cells. Scale bar: 10μm. (D) [¹⁴C]-Oleate oxidation assay confirms etomoxir inhibited FAO activity in cultured NSCs. (E and F) Etomoxir potentiated NSC differentiation in organotypic culture of forebrain hemispheres. (E) Representative images from three independent experiments showing reduced thickness of ventricular zone (Pax6⁺ layer) in etomoxir group and increased fraction of Pax6⁺ cells that co-express Tbr2. The pial surface is outlined by dashed lines. Scale bars: 20μm. (F) Quantifications. The percentage of Pax6⁺ cells that co-express Tbr2 was significantly increased in the etomoxir group.

(G and H) Blocking fatty acid mobilization from lipid droplets by overexpressing dominant-negative mutants of PLIN1 diminished the NSC pool. Mouse embryos were electroporated at E12.5 and sacrificed at E15.5. (G) Representative confocal images. Areas of the ventricular zone (Pax6⁺ layer) are shown at higher magnification in bottom panels. Scale bars: 50μm. (H) Quantification and statistics. See also Figure S3.