Fig. 1

Analysis of gliosis marker. a-d NPC1 mutant cell lines contained a higher amount of GFAP⁺ and vimentin⁺ cells (red, a-d). DAPI staining (blue) indicates nuclei. Scale 100 μm. (e). Colocalization analysis of GFAP and vimentin revealed a significantly increased amount of double positive cells in all NPC1 mutant cell lines. f FACS analysis of GFAP⁺/vimentin⁺ cells confirmed an increased amount of glia cells in NPC1 mutant cell lines (N = 5–7, n = 14–23). g In addition, semi-quantitative protein measurement by western blot demonstrated a higher amount of GFAP (N = 7–11, n = 26–43) and h an increased amount of vimentin in NPC1 mutant cells (N = 5–7, n = 14–23). i FACS analysis of cells positive for GFAP and Ki67, elucidated an increased amount of double positive cells indicating gliosis NPC1 mutant cell lines (g; N = 3–4, n = 7–9)