Figure 4 –

Zafirlukast effects downstream measures of thiol isomerase inhibition. (A) Zafirlukast treatment inhibits the activation of the EGFR receptor (n=3). Data is presented as mean ±SD. A student’s t-test where *p=0.0112 for 1 hour 30 µM treated zafirlukast and ***p= 0.0002 for 4 hour 30 µM treated zafirlukast compared to the EGF control. (B) The known ERp57 inhibitor RL90 and PACMA 31 also inhibit EGFR activation (n=3). Data is presented as mean ±SD. A student’s t-test where ***p=0.0007 for 30 µM zafirlukast, **p=0.004 for 5 µg RL90, *p=0.0433 for 10 µg RL90, **p=0.0059 for 15 µg RL90 and **p=0.0092 for 10 µM PACMA 31 compared to the EGF control. (C) A time-course study of zafirlukast RL-90, and PACMA 31’s effect on OVCAR8 cell viability (n=4). Data is presented as mean ±SD. A two-way ANOVA and a post-hoc Sidak’s test demonstrate no significant changes between the control and any of the test groups. (D) Phosphorylation of Gab1, an immediate downstream target of EGFR phosphorylation (n=3) Data is presented as mean ±SD. A student’s t-test where *p=0.0153 for 1 hour 10 µM zafirlukast and ****p<0.0001 for 4 hour 10 µM and 1 and 4 hour 30 µM zafirlukast compared to the EGF control. (E) Zafirlukast inhibits the tissue factor dependent generation of Factor Xa (n=4). Data is presented as mean ±SD. A one-way ANOVA and post-hoc Dunnett’s test where **p=0.0041 for 10 µM zafirlukast and ****p<0.0001 for 30 µM zafirlukast.