Activation of ECs by anti-β₂GPI Abs induces increased expression of components of the multiprotein signaling complex and accessory proteins. (A) ECs were incubated with β₂GPI and anti-β₂GPI Abs or TNFα to induce activation. Total RNA was then isolated, reverse-transcribed, and analyzed for content of S100A10 and annexin A2 mRNA using qPCR. Significant increases in both mRNA species occurred in response to β₂GPI and anti-β₂GPI Abs, but not TNFα. (B) ECs were activated as described in panel A, and RNA isolated, reverse transcribed, and analyzed for content of MyD88, MD2, TLR4, calreticulin, and nucleolin mRNA using qPCR. Significant increases in mRNA species encoding MyD88 and MD2, with lesser increases in mRNA encoding TLR4, calreticulin, and nucleolin, were observed. These exceeded those observed in response to TNFα. Error bars represent the means ± SD of triplicate points.