Fig. 5
From: Axl receptor tyrosine kinase is a regulator of apolipoprotein E
AZ7235 has nominal effects on secretion and mRNA levels of apoE and no effect on ABCA1 expression in primary murine mixed glia expressing human apoE3. Primary mixed glial cells (~ 90% astrocytes, 10% microglia) were cultivated from neonatal human APOE3 targeted-replacement mice. Cells were re-plated at 18–20 days in vitro followed by treatment with vehicle (DMSO), 1 μM T0901317, 1 μM and 3 μM AZ7235 for 72 h. (a) Secreted apoE levels in conditioned media were measured by apoE ELISA. The graph represents mean concentration and standard deviation from 4 independent experiments. (b) Representative immunoblot of cellular ABCA1 and apoE with GAPDH as the loading control. Immunoblot images were cropped to show relevant lanes. (c) Quantification of cellular apoE and (d) ABCA1 protein levels in murine glia lysates by immunoblotting. (e-f) Human APOE (e) and mouse Abca1 (f) mRNA levels were measured by qRT-PCR. Graphs represent fold-change over DMSO control (dashed line) and +/− 95% confidence intervals from N independent experiments indicated in brackets. ** P < 0.01, *** P < 0.001 compared to vehicle control by blocked ANOVA post-hoc tests