Fig. 2.

Effect of EROS knockdown on ERK1/2 phosphorylation and abundance of EROS, NOX2 and RAC1

A. Representative immunoblot shows ERK1/2 phosphorylation in response to histamine and VEGF stimulation in siControl transfected HUVEC, which is strongly attenuated in EROS knockdown cells. B. Statistical analysis of ERK1/2 phosphorylation (n = 3 for all conditions). C. Quantitative RT-PCR of HUVEC mRNA downregulating EROS reveals low EROS (yellow bar, n = 6), unchanged RAC1 (blue bar, n = 6) as well as NOX4 (gray bar, n = 6) and high NOX2 transcripts (red bar, n = 6). D. Western blots of protein lysates from HUVEC transfected with either siControl, siRAC1, siEROS or siNOX2 probed with antibodies for NOX2, RAC1, EROS and GAPDH. E-G. EROS, NOX2 and RAC1 are significantly downregulated within all single-guided siRNA samples, siEROS (yellow bars), siNOX2 (red bars) and siRAC1 (blue bars) compared to siControl (green bars). All values are presented as means ± SEM, **P < 0.01, ***P < 0.001 and ****P < 0.0001 compared to untreated siControl HUVEC, ^#P < 0.05 of EROS siRNA vs. NOX2 siRNA and ^##P < 0.01 compared to same treatments, either histamine or VEGF, following siRNA knockdown of RAC1 vs. Control siRNA using 1-way ANOVA. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)