Research Article
A Fragment of Apolipoprotein E4 Leads to the Downregulation of a CXorf56 Homologue, a Novel ER-Associated Protein, and Activation of BV2 Microglial Cells
Figure 5
Treatment of BV2 microglial cells with an amino-terminal fragment of apoE4 leads to the downregulation of CXorf56. (a) BV2 cells were treated for 5 hours with 25 μg/mg nApoE41-151 (blue bar) or with an equivalent concentration of full-length apoE4 (red bar). Following treatment, RNA was extracted and RT qPCR was performed. Data indicated a significant decrease in the expression of the CXorf56 homologue following treatment, , while full-length apoE4 had no significant effect. (b, c) Western blot analysis utilizing an anti-CXorf56 antibody (1 : 500) confirmed the decreased expression of the CXorf56 protein following treatment of BV2 cells for 24 hrs with 25 μg/ml nApoE41-151. Beta-actin was employed as a loading control utilizing an anti-beta-actin antibody (1 : 50K), bottom panel. (c) Quantitative densitometry analysis of 3 independent experiments showing an overall 74% decrease in the expression of CXorf56 protein following treatment ( denotes ). Data are representative of 3 independent (d, e) BV2 cells were left untreated (d) or treated for 24 hours with nApoE41-151 and stained with the CXorf56 antibody. Following treatment, there was a decrease in the staining intensity as well as a morphological shift to bipolar, extended pseudopods (arrowheads, e).
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