Mab-10H10, but not Mab-5G9, dissociates the TF-α3β1 integrin complex in HaCaT cells. (A) TF associates with α3 and α6 in HaCaT cells. VIIa was added for 60 minutes, and lysates were immunoprecipitated with anti-β1, α2, α3, and α6. β1 was used as a control for the efficiency of immunoprecipitation of integrin heterodimers, and coprecipitation of TF was assessed by Western blotting. Immunoprecipitation of α5 yielded no signal, consistent with the low expression levels of this integrin subunit in HaCaT cells.¹⁷  Densitometric quantitation is given as mean and standard deviation, n = 3. (B) TF associates with α3 and α6 in endothelial cells. HUVECs were transduced to express TF and PAR2. VIIa was added for 60 minutes, and lysates were immunoprecipitated with anti-α2, α3, α5, and α6. The predepleted lysates were then reimmunoprecipitated with anti-β1. Coprecipitation of TF was assessed by Western blotting. β1 was assessed on blot as well as a control for the efficiency of immunoprecipitation of the β1 subunit as well as integrin heterodimers. (C) Mab-5G9 stimulates and Mab-10H10 inhibits TF association with α3 and β1 integrin in HaCaT cells. Cells were incubated for 60 minutes with 50 μg/mL Mab-10H10 or Mab-5G9 and integrin β1 and α3 were immunoprecipitated from lysates. β1 and TF in immunoprecipitates and supernatants were determined by Western blot. The graph shows mean and standard deviation for 3 β1 pulldown assays. (D) Mab-10H10 inhibits basal and VIIa-induced TF association with β1 integrin. HaCaT cells were incubated with 50 μg/mL Mab-10H10 or Mab-5G9 for 15 minutes and treated with VIIa for 60 minutes.