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

Figure 4.

Figure 4

Crabp2 promotes integrin β1/FAK/ERK signaling via HuR. (a) C10F4 cell lysate was immunoprecipitated with Crabp2 antibody or rabbit IgG (as control), respectively. Western blot analysis was performed to detect Crabp2 and HuR in input lysate (Input) and co-immunoprecipitated proteins. (b and d) Real-time RT-PCR of HuR in C10F4 cells expressing control-shRNA (shCon) versus Crabp2-shRNA (shCrabp2) (b), or C10F4 cells expressing empty vector versus Crabp2 (d). The p values were determined using Student’s t test. (c,e) Real-time RT-PCR of integrin β1 (Itgb1) in C10F4 cells expressing control-shRNA or Crabp2-shRNA (c), or C10F4 cells expressing Vector + shCon, Crabp2 + shCon, or Crabp2 + shHuR (e). The p values were determined using Student’s t test (c) or one-way ANOVA (e). **p < 0.01, ***p < 0.001. (f) Western blot analysis of HuR, integrin β1 (ITGB1), phosphorylated FAK (Y397), total FAK, phosphorylated ERK1/2 (T202/Y204), total ERK1/2, Crabp2, and β-actin (ACTB, as the internal control) levels in C10F4 cells expressing control-shRNA or Crabp2-shRNA. (g) Western blot analysis of integrin β1, phosphorylated ERK1/2 (T202/Y204), total ERK1/2, CRABP2, and β-actin (ACTB, as the internal control) levels in H1650 cells expressing control-siRNA or CRABP2-siRNA. (h) Western blot analysis of HuR, Crabp2, and β-actin (ACTB, as the internal control) in C10F4 cells expressing empty vector or Crabp2. (i) Western blot analysis of integrin β1 (ITGB1), phosphorylated FAK (Y397), total FAK, phosphorylated ERK1/2 (T202/Y204), total ERK1/2, and β-actin (ACTB, as the internal control) levels in C10F4 cells expressing Vector + shCon, Crabp2 + shCon, or Crabp2 + shHuR.