Fig. 6.
In HUVECs, apabetalone regulation of transcription reduces the abundance of VCAM-1 and MCP-1 proteins. HUVEC cells were stimulated with TNFα and co-treated with apabetalone for 4 h. The surface abundance of VCAM-1 (FITC-CD106) and SELE (APC-CD62E) were measured by flow cytometry. a Representative histogram overlay of HUVEC surface staining for VCAM-1 and SELE. Smaller peaks (% positive reduction) and left-ward curve shifts (MFI reduction) are both indications that there is a reduction in surface expression for the given protein. b Average of % positive cells expressing VCAM1 or SELE on the cell surface relative to the isotype control (the filled gray histogram as indicated in A). c Average mean fluorescent intensity (MFI) of VCAM1 and SELE on HUVEC surface relative to DMSO control. d HUVEC MCP-1 secretion is induced by overnight TNFα stimulation. Co-application with 20 μM apabetalone significantly reduces MCP-1 secretion (BDTM cytometric bead array). In b–d, the results represent the mean of four independent experiments ± standard error. Statistical significance was determined through 1-way ANOVA analysis followed by Dunnett’s Multiple Comparison Test using TNFα response for the comparison, where *p < 0.05, **p < 0.01, ***p < 0.001