Fig. 4

The effect of ATRA treatment on expression of ANXA2 and S100A10 mRNA and protein levels. The effect on ANXA2 and S100A10 expression following 1–6 days ATRA treatment (1, 5 μM) in OAW28 (a and c) and OV-90 (b and d) cells. ANXA2 and S100A10 expression was assessed using 2^-∆∆CT quantitation method and normalised to housekeeping gene β-actin using no treatment control as a calibrator. Data is from 2 to 4 independent experiments (n = 4–12). Statistical significance was determined using one-way ANOVA and the Tukey multiple comparison post hoc test. Western blots for annexin A2 and S100A10 (e and f) following 6 days ATRA treatment (1, 5 μM). Annexin A2 bands at 38 kDa, S100A10 at 11 kDa and β-actin bands at 42 kDa are shown for OAW28 (e) and OV-90 (f) cells. Equal amounts protein (5 μg for annexin A2 & 15 μg S100A10 were run on a 4–12% SDS-PAGE gel and immunoblotted with mouse monoclonal antibodies to annexin A2 (1/2000, BD Biosciences) or S100A10 (1/2000, BD Biosciences). A rabbit polyclonal antibody was used detect β-actin (1/2000, ab8227, Abcam). Numbers below western bands in e and f are fold changes relative to the control treatment. Westerns blots are representative of two independent experiments