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. 2014 Nov 28;3(12):1228–1235. doi: 10.1242/bio.201410132

Fig. 2. Impaired β1 integrin activation correlates with the inability of EC to respond to fluid shear stress.

Fig. 2.

(A) HUVEC and EA.hy926 monolayers after static condition or fluid shear stress at 2.0 Pa for 18 h. White arrow: indicates the direction of flow. Cells were stained by activated-state-specific β1-integrin antibody (Clone:9EG7). (B) Percentage of activated β1 integrin in the total β1 integrin population on the surface of HUVEC and EA.hy926 cell line analyzed by flow cytometry. Data are means ± SE (N = 3), *p<0.05. (C) Silencing of β1 integrin by siRNA in HUVEC analyzed by western blot. (D) CT- or β1 integrin-silenced HUVEC were maintained under static conditions or exposed to fluid shear stress at 2.0 Pa for 5 h. F-actin and activated β1 integrin were stained. White arrow: indicates the direction of flow. (E) Actin stress fiber orientation (0° indicates perfect alignment with flow direction; 45° indicates random orientation). Data are means ± SE (N = 6), *p<0.05. CT- or β1 integrin-silenced HUVEC exposed to fluid shear stress at 2.0 Pa for 5 h were stained by (F) VE-cadherin and activated-state-specific β1-integrin antibody (Clone:9EG7) and (G) by paxillin antibody. Scale bars: 10 µm (A), 50 µm (D), 20 µm (F,G).