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. Author manuscript; available in PMC: 2010 Jul 21.
Published in final edited form as: Oncogene. 2009 Nov 9;29(3):368–379. doi: 10.1038/onc.2009.360

Figure 3. Reducing EYA4 expression with shRNA inhibits cell adhesion and migration.

Figure 3

(a) EYA4 protein is expressed in MPNST cell lines (STS26T, ST8814, S462, T265) but not normal human Schwann cells (NHSC), assessed by western blotting. Staining with anti-actin confirmed equal amounts of protein across samples. (b, c) EYA4 expression in S462TY MPNST cells is reduced subsequent to introduction of shEYA4 RNAs (4.3, 4.4, 4.5, 4.7). Data are shown for level of RNA using QRT-PCR (b) and protein by western blotting with anti-EYA4 or an anti-AKT loading control (c). (d) Phase contrast micrographs of living S462TY cells at day 0 (d0) and day 4 (d4). Fewer S462TY cells containing shEYA4 shRNAs attach to dishes, correlating with decreased cell numbers at day 4. (e) Adhesion assay of S462TY cells containing control or shEYA4 cells. Significantly fewer cells remain attached (*p<0.05) in the presence of EYA4 shRNAs (4.3, 4.7) relative to control shRNA. (f) Migration of S462TY MPNST cells through transwell filters is significantly reduced (*p<0.05; **p<0.01) in the presence of EYA4 shRNAs (4.3, 4.4, 4.5). Values are expressed as percent migration relative to control shRNA.