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. 2016 Feb 11;7(12):14350–14365. doi: 10.18632/oncotarget.7320

Figure 2. LDE225 inhibits the VLA4-mediated FAK signaling pathway in MCL cells and the production of IL-6, SDF-1 and VCAM-1 in stromal cells.

Figure 2

A. The mRNA levels (Left) and protein levels (Right) of transducers in the FAK signaling pathway were measured by qRT-PCR and immunoblots. SP53 MCL cells and primary cells were treated with LDE225 (30 μM) or DMSO. Each value in qRT-PCR was normalized to GAPDH and represents the mean ± S.D. from three independent experiments. The protein levels were analyzed using of a Gel-Pro Analysis software from three independent immunoblots, and GAPDH was used as a loading control. B. Mean fluorescence intensity (MFI) of VLA-4 in MCL cell lines and patient samples were decreased after treatment of LDE225 (10 μM or 30 μM) compared with DMSO in a dose-dependent manner. C. Immunoblot analyses of IL-6 (Top Left) and SDF-1 (Top Right) using conditioned media from HS5 and HS27a stromal cells treated with LDE225 (30 μM). Conditioned media from HS5 and HS27a cells treated with DMSO was used as controls. FACS analysis showed reduced expression of VCAM-1 in HS5 and HS27a cells treated with LDE225. Each value in the diagram was normalized to MFI of VCAM-1 in the cells treated with DMSO (Bottom). D. MCL cells were stained with PKH26 and were seeded onto the monolayer of HS5 or HS27a BMSCs, which were pre-treated with LDE225 treatment (30 μM) or DMSO for 72 h. PKH26 dye intensity was analyzed and shown as the mean ± S.D. from three independent experiments. NS, not significant,*p < 0.05, **p < 0.01 (vs. cells treated with DMSO; Student's t-test).