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Fig. 7 | Acta Neuropathologica Communications

Fig. 7

From: Impaired lipid metabolism in astrocytes underlies degeneration of cortical projection neurons in hereditary spastic paraplegia

Fig. 7

GW3965 treatment increases cholesterol content of SPG3A cortical PNs by promoting cholesterol efflux from astroglial cells in mutant ATL1 neural cultures. a Representative fluorescence and corresponding phase images of Filipin staining of ATL1-P342S and ATL1-A161P #70 cortical PNs after GW3965 or DMSO treatment for 3 days at 10 weeks in culture. Scale bar: 20 µm. b, c Quantification of Filipin staining intensity in cell bodies (b) and axons (c) of GW3965-treated ATL1-P342S and ATL1-A161P #70 cortical PNs. d Cholesterol efflux in ATL1-P342S and ATL1-A161P #70 regular neural cultures after GW3965 or DMSO treatment for 3 days. e Immunostaining of astroglial cell makers, GFAP in ATL1-P342S and ATL1-A161P #70 astroglial cells. Red: GFAP, cyan: Hoechst. Scale bar: 50 µm. f NR1H2 and APOE mRNA levels in WT, ATL1-P342S, ATL1-342-Cor, H9 and ATL1-A161P #70 astroglial cells. Data are represented as mean ± SEM. *p < 0.05 compared to WT (for ATL1-P342S) and H9 (for ATL1-A161P #70). #p < 0.05 compared to ATL1-P342S. g Cholesterol efflux from ATL1-P342S and ATL1-A161P #70 enriched cortical PNs at D49 after 1 µM GW3965 or DMSO treatment for 3 days. h Cholesterol efflux from ATL1-P342S and ATL1-A161P #70 astroglial cells after 1 µM GW3965 or DMSO treatment for 3 days. i Cholesterol efflux-associated gene expression in ATL1-P342S and ATL1-A161P #70 astroglial cells after GW3965 (1 µM) or DMSO treatment for 3 days. Data are represented as mean ± SEM. *p < 0.05 compared to DMSO treated ATL1-P342S and ATL1-A161P #70 astroglial cells, respectively, by two-sided Student’s t-test

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