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. 2021 Jan 12;11:644. doi: 10.1038/s41598-020-80060-3

Figure 2.

Figure 2

EPHA10 influences cell migration, epithelial–mesenchymal transition, tumorsphere formation and gene expression in LN1-1 cells. (A) Left: Representative images of migrated cells. Scale bars, 100 μm. Right: Relative migration activity of the EPHA10 knockdown cells was calculated by normalizing the mean number of migrated cells per field (EPHA10 sh3 and sh5, n = 10) to that of the control cells (pLKO-GFP, n = 10). (B) Morphology of LN1-1 pLKO-GFP, EPHA10 sh3, and EPHA10 sh5 cells. Scale bars, 20 μm. (C) Immunoblot analysis of α-catenin, β-catenin, vimentin, and E-cadherin proteins in LN1-1 cells with EPHA10 knockdown (EPHA10 sh1–5) and the control vector (pLKO-GFP). Protein levels were normalized to α-tubulin. Relative ratios were determined by dividing the level of the protein of interest in each expression variant by that in the pLKO-GFP vector-expressing cells. (D) Left: Representative images of tumorspheres in LN1-1 pLKO-GFP, EPHA10 sh3, and EPHA10 sh5 cells. Scale bars, 100 μm. Right: Relative sphere formation activity was determined by normalizing the mean number of spheres per field for the EPHA10 sh3 and sh5 cells (n = 2) to that of the pLKO-GFP cells (n = 2). (E) The levels of TWIST, SNAIL, SLUG, OCT4, NANOG, and SOX2 mRNA in LN1-1 pLKO-GFP, EPHA10 sh3, and EPHA10 sh5 cells were determined by qRT-PCR. The amplifications were first normalized to β-actin (internal control). For each gene, the relative expression in LN1-1 EPHA10 sh3 and EPHA10 sh5 cells (n = 3) was normalized to that in the LN1-1 pLKO-GFP cells (n = 3). Bars represent SE; *p < 0.05; **p < 0.01; ***p < 0.001.