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. 2014 Jan 13;4:3652. doi: 10.1038/srep03652

Figure 4. SCRG1/BST1 stimulates FAK/PI3K-dependent hMSC migration and preserves migratory activity after ex vivo expansion.

Figure 4

(a) Migration of UE7T-13 cells was investigated as described in the Methods. rhSCRG1 was added at various concentrations (1–1000 ng/mL). After incubation for 15 h, the number of cells that had migrated to the lower side was counted. (b and c) UE7T-13 cells were transfected with siSCRG1 (b) or siBST1 (c). rhSCRG1 (500 ng/mL) was added to the lower well of the Transwell plate and trans-well migration was analyzed as described in (a). (d) UE7T-13 cells were transfected with pCMV-null-IRES-AcGFP (pCTRL) or pCMV-BST1-IRES-AcGFP (pBST1). rhSCRG1 (500 ng/mL) was added to the lower well of the Transwell plate and trans-well migration was analyzed as described in (a). (e) Phosphorylation of FAK was detected in UE7T-13 as in Figs. 2a–c. Although cropped blots were used, the gels were run under the same experimental conditions. (f) Phosphorylation of FAK was detected in UE7T-13 as in Fig. 3b. Although cropped blots were used, the gels were run under the same experimental conditions. (g) UE7T-13 cells were treated with 1 μM kinase inhibitors trans-well migration was analyzed as in (a). PI3K inhibitor LY294002 (PI3K), MEK inhibitor U0126 (MEK), JNK inhibitor SP600125 (JNK), and FAK inhibitor I (FAK) were added to the lower and upper wells. (h) Primary cultured hMSCs (passage number #5) were subcultured ten times in the presence (passage number #15, +rhSCRG1) or absence (passage number #15, −rhSCRG1) of 500 ng/mL rhSCRG1. Trans-well migration was analyzed as in (a). In a–d, g, and h, data are presented as mean ± SD. *p < 0.05 was considered significant.