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Fig. 4 | BMC Cancer

Fig. 4

From: The effect of soluble E-selectin on tumor progression and metastasis

Fig. 4

Soluble E-selectin triggers FAK activation: a Activation of FAK by sE-selectin in MDA-MB-231. An MDA-MB-231 suspension was incubated with sE-selectin (100 nM) for the indicated period of time (0-60 min). CD44 shRNA knockdown clones or control shRNA of MDA-MB-231 cells (clone 28) were incubated with sE-selectin for the indicated time. Cell lysate (30 μg) was separated by SDS-PAGE and probed with pFAK (Y397) and total FAK, followed by HRP-conjugated secondary antibody. GAPDH was used as a loading control. The blot was visualized by ECL. b FAK inhibitor blocks sE-selectin-mediated adhesion. MDA-MB-231 or HL-60 cells were co-incubated with vehicle alone, sE-selectin, FAK inhibitor II (5 μM), or sE-selectin + FAK inhibitor II for 30 min in a rotation wheel. After a brief wash with PBS, the cells were infused into parallel flow chambers over a 5 min period at a shear stress rate of 1 dyn/cm2. The adherent cells were counted under a microscope. The data were summarized as fold increase of adhesion of MDA-MB-231 and HL60 cells. The data represent Mean ± SD. Statistical significance was determined by Student’s t-test

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