Fig. 1

Increased lysosomal activity in qNSCs in vitro. a Peptidase activities in NSCs. Trypsin-like, chymotrypsin-like, and caspase-like activities in NSC lysate were continuously measured every 5 min for 1 h, with or without proteasome inhibitor (PI) or cathepsin inhibitor (CI); n = 2. b qPCR of the lysosomal genes: Lamp1 and cathepsins (CtsA, CtsB, and CtsF), and the transcriptional regulator TFEB, after BMP exposure. c Immunocytochemistry of Lamp1 (red) with DAPI (blue) (upper panels). Staining of Magic Red cathepsin L (red) and Hoechst 33342 (blue) (lower panels). Scale bars, 50 µm. d, e Immunocytochemistry of Lamp1 and TFEB in the active and quiescent states induced by BMP in NSCs, NS5 cells (d) and adult NSCs (e). Selected regions indicated by white squares are enlarged. Yellow arrowheads in enlarged squares indicate nuclear localization of TFEB in the quiescent state. Scale bars, 100 µm. f Lysosomal cathepsin L activity in NSCs, as determined using Magid Rred cathepsin L; n = 3. Corresponding data points were plotted. g Immunoblots of signaling molecules in NSCs after BMP exposure. β-Actin was used as a loading control. h Immunoblotting of several NSC lines in the active and quiescent states. Phosphorylated (##) and dephosphorylated forms of TFEB (#) were detected. i Reporter assay of TFEB-promoter–driven luciferase; n = 4. Data represent means ± s.e.m. (**P < 0.01; Student’s t-test). Source data of immunoblots are provided as a Source Data file.