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. 2019 Jan 29;9:845. doi: 10.1038/s41598-018-37443-4

Figure 5.

Figure 5

Crabp2 promotes migration, invasion, and anoikis resistance via HuR and integrin β1/FAK/ERK signaling. (a) Western blot analysis of HuR of C10F4 cells expressing control-siRNA or HuR-siRNA (A, B, or C). (b) Western blot analysis of integrin β1 (ITGB1) of C10F4 cells expressing control-siRNA or ITGB1-siRNA (A, B, or C). (c) Migration (up) or invasion (down) assay of C10F4 cells expressing Vector + siCon, Crabp2 + siCon, Crabp2 + siHuR (siHuR-C), Crabp2 + siITGB1 (siITGB1-A), Crabp2 + FR180204 (40 µM), or Crabp2 + FAK inhibitor 14 (1 µM). Cells migrated into the lower compartment of Boyden chamber were photographed (left) and quantified (right). The p values were determined using one-way ANOVA. (d,e) Anoikis assay or C10F4 cells expressing Vector + siCon, Crabp2 + siCon, or Crabp2 + siHuR (siHuR-C) (d), or C10F4 cells expressing Vector + siCon, Crabp2 + siCon, or Crabp2 + siITGB1-A/C (e). Cells were plated onto a 96-well anchorage-resistant plate at a density of 10,000 cells/well. Twenty-four hours later, viability of cells was assessed by MTS assay. The p values were determined using one-way ANOVA. (f,g) C10F4 cells expressing empty vector or Crabp2 were plated onto a 96-well anchorage-resistant plate at a density of 10,000 cells/well and treated with DMSO (as control), ERK1/2 inhibitor FR180204 (ERKi, 40 or 80 μM) (f), or FAK inhibitor 14 (FAKi14, 1 μM) (g). Twenty-four hours later, viability of cells was assessed by MTS assay. The p values were determined using one-way ANOVA *p < 0.05, **p < 0.01, ***p < 0.001.