American Society of Hematology

Figure 6.

$Effect of RUNX1 KD on αIIbβ3 expression and activation, MLC expression and phosphorylation, and IFITM3 in HEL cells. (A) Effect of RUNX1 KD on αIIbβ3 expression in HEL cells by flow cytometry using anti-CD41a antibody. (B) Representative immunoblots of αIIb, β3, RUNX1, and β-actin protein expression and densitometric quantification (n = 4). P values show comparisons by the Student t test. (C) Effect of RUNX1 KD on αIIbβ3 activation evaluated by flow cytometry. FITC-labeled PAC1 antibody was used to assess activation of αIIbβ3 complex in control HEL cells (black lines) and after RUNX1 KD (red lines), in the resting state (interrupted lines) and upon stimulation with ADP (50 μM) or thrombin (10 U/mL) (continuous lines). Data are expressed in arbitrary units as mean ± SEM (n = 3). (D) Effect of RUNX1 KD on MLC expression and its phosphorylation upon thrombin activation in HEL cells. Left panel: representative immunoblots of total MLC, phospho-MLC (pMLC), RUNX1, and actin protein expression in control cells and those with RUNX1 KD, in the resting state (baseline) and after thrombin activation (5 U/mL). Right panel: relative protein expression of RUNX1 and total MLC, and of pMLC in resting state (baseline) and after thrombin (5 U/mL) activation in control and RUNX1-deficient cells. The last 2 bars show the pMLC as a fraction of the total MLC present in control cells and RUNX1 deficient cells. (E) Representative immunoblots showing relative protein expression in HEL cells of IFITM3, RUNX1, and actin, and densitometric quantification on RUNX1 KD. Data are shown as mean ± SEM (n = 4).$

Effect of RUNX1 KD on αIIbβ3 expression and activation, MLC expression and phosphorylation, and IFITM3 in HEL cells. (A) Effect of RUNX1 KD on αIIbβ3 expression in HEL cells by flow cytometry using anti-CD41a antibody. (B) Representative immunoblots of αIIb, β3, RUNX1, and β-actin protein expression and densitometric quantification (n = 4). P values show comparisons by the Student t test. (C) Effect of RUNX1 KD on αIIbβ3 activation evaluated by flow cytometry. FITC-labeled PAC1 antibody was used to assess activation of αIIbβ3 complex in control HEL cells (black lines) and after RUNX1 KD (red lines), in the resting state (interrupted lines) and upon stimulation with ADP (50 μM) or thrombin (10 U/mL) (continuous lines). Data are expressed in arbitrary units as mean ± SEM (n = 3). (D) Effect of RUNX1 KD on MLC expression and its phosphorylation upon thrombin activation in HEL cells. Left panel: representative immunoblots of total MLC, phospho-MLC (pMLC), RUNX1, and actin protein expression in control cells and those with RUNX1 KD, in the resting state (baseline) and after thrombin activation (5 U/mL). Right panel: relative protein expression of RUNX1 and total MLC, and of pMLC in resting state (baseline) and after thrombin (5 U/mL) activation in control and RUNX1-deficient cells. The last 2 bars show the pMLC as a fraction of the total MLC present in control cells and RUNX1 deficient cells. (E) Representative immunoblots showing relative protein expression in HEL cells of IFITM3, RUNX1, and actin, and densitometric quantification on RUNX1 KD. Data are shown as mean ± SEM (n = 4).

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