Fig. 1

AZ7235 increases apoE expression in various CNS cell types. (a-b) CCF-STTG1 cells were treated with AZ7235 using a 7-point concentration response curve at the indicated concentration (0.041–30 μM) for 72 h. (a) ApoE secretion was measured by ELISA and data are expressed as % apoE secretion relative to DMSO (0%) and 1 μM of the positive control LXR agonist T0901317 (100%). (b) Cell viability was measured by CellTiter-Blue assay and data are expressed as percentage change relative to DMSO treatment (100%, dashed line). (c) Log dose response curves for other Axl inhibitors R428, S49076 and UNC2025 (0.1 μM – 3 μM) on apoE secretion in CCF-STTG1 after 72 h treatment. Data are expressed as fold-change relative to DMSO treatment (dashed line). Error bars represent range of duplicate wells in one representative assay. (d) APOE mRNA levels were measured by qRT-PCR and (e) cellular apoE protein levels were measured by immunoblot in CCF-STTG1 cells after 72 h treatment with vehicle control DMSO, 1 μM T0901317, 3 μM AZ7235. (f) Representative immunoblot of cellular apoE. Images were cropped to show relevant lanes. Graphs of (d) and (e) represent fold-change over DMSO control (dashed line) and +/− 95% confidence intervals from N independent experiments indicated in brackets. (g-i) Secreted apoE levels were measured in primary human astrocytes (g), HMC3 (h) and primary human brain vascular pericytes (i) after 72 h drug treatment. Graph represents mean concentration and standard deviation from N independent experiments indicated in brackets. ** P < 0.01, *** P < 0.001 compared to vehicle control using blocked two-way analysis of variance (ANOVA) post-hoc tests