Figure 3.
aPLs induce thrombin-dependent NADPH oxidase endosomal internalization by the TF-FVIIa-integrin complex in human monocytes. (A) Live cell imaging of HL5B (green) internalization after 15 minutes and HL5B-mediated NOX2 (gp91phox, green) trafficking into the endosome after 30 minutes in the presence of indicated inhibitors. Nuclei were stained in blue with Hoechst 33342 or 4′,6-diamidino-2-phenylindole, surface membrane with cholera toxin B (CTB), endo-lysosomes with Lysotracker or αEEA1 (red). Scale bar = 10 µm (upper panels) or 5 µm (lower panel). (B) TF and TNFα mRNA expression and ROS production by MM1 cells treated as indicated; mean ± SD. n = 6. *P < .0001; 1-way ANOVA, Dunnett multiple-comparison test compared with control sample. (C) Schematic representation of aPL-induced PAR1/PAR2 heterodimer signaling linked to TF-VIIa-integrin complex trafficking. (D) Dose response of rivaroxaban inhibition of aPL HL5B-induced TNFα expression by MM1 cells.

aPLs induce thrombin-dependent NADPH oxidase endosomal internalization by the TF-FVIIa-integrin complex in human monocytes. (A) Live cell imaging of HL5B (green) internalization after 15 minutes and HL5B-mediated NOX2 (gp91phox, green) trafficking into the endosome after 30 minutes in the presence of indicated inhibitors. Nuclei were stained in blue with Hoechst 33342 or 4′,6-diamidino-2-phenylindole, surface membrane with cholera toxin B (CTB), endo-lysosomes with Lysotracker or αEEA1 (red). Scale bar = 10 µm (upper panels) or 5 µm (lower panel). (B) TF and TNFα mRNA expression and ROS production by MM1 cells treated as indicated; mean ± SD. n = 6. *P < .0001; 1-way ANOVA, Dunnett multiple-comparison test compared with control sample. (C) Schematic representation of aPL-induced PAR1/PAR2 heterodimer signaling linked to TF-VIIa-integrin complex trafficking. (D) Dose response of rivaroxaban inhibition of aPL HL5B-induced TNFα expression by MM1 cells.

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