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. 2015 Apr 17;10(4):e0123432. doi: 10.1371/journal.pone.0123432

Fig 7. Orai1 knockdown decreases HMGB1-induced permeability.

Fig 7

A. Orai1 protein expression after RNA inference. EA.hy926 cells were transfected for 48 h with Orai1 siRNA-1, Orai1 siRNA-2 or control (scrambled) siRNA. Cells were harvested and total protein was extracted and subjected to western blotting with anti-Orai1 antibodies, with anti-GAPDH antibodies as a loading control. Orai1 expression was quantified and analyzed statistically based on three independent experiments. B. HMGB1-induced permeability was inhibited by Orai1 knockdown. EA.hy926 cells were plated in the upper part of transwell chambers until the formation of a tight monolayer, then transfected with Orai1 siRNA-1, Orai1 siRNA-2 or control (scrambled) siRNA. HMGB1 200 ng/ml was added and cells were incubated for an additional 24 h. After incubation, endothelial permeability was assessed, as described above. Data are presented as mean ± SD of three independent experiments. *Indicates significant difference compared with the control group (P<0.05).