Fig. 2

Lysosomal inhibition to qNSCs. a Bright field view of NSCs; bipolar proliferating NSCs (aNSC), flat qNSCs (qNSC), and bipolar qNSCs treated with 20 nM bafilomycin A (qNSC, BafA). b Immunocytochemistry of Ki-67. Representative view of NSCs immunostained for Ki-67 (green) and Sox2 (red). qNSCs were incubated in proliferation medium containing EGF (qNSC, PM), quiescence medium (qNSC), or quiescence medium containing BafA (qNSC, BafA) for 1 day, and then analyzed. c Percentage of Ki-67–positive (Ki) or EdU-incorporated (EdU) cells. Results are averages of eight samples of qNSCs (control), BafA-treated qNSCs (BafA), or qNSCs after incubation with proliferation medium (PM); n = 8. d Immunoblotting of qNSCs after incubation with qNSC medium (control) or qNSC medium containing BafA (20 nM) (BafA) for 4–20 h. e Immunocytochemistry of EGFR. BafA-treated qNSCs accumulated EGFR (green) in the cytoplasm. EGFR on the cell surface was not detected by immunostaining following membrane permeabilization. This might be responsible for the differences between the results of immunostainings and western blotting. f Effect of medium change. Samples on the left sides of blots were incubated in fresh quiescence medium, with or without BafA. In the w/o medium change samples, the medium in culture dishes was partially removed and added back with or without addition of BafA. Scale bars, 100 µm. Data represent means ± s.e.m. (*****P < 0.0001; Student’s t-test). Source data of immunoblots are provided as a Source Data file.