Figure 3.
YAP mediates TNFα-induced expression of HK2 and cell migration in breast cancer cells. (a, b) Control and shYAP MCF7 cells were treated with 10 μM TNFα for 12 h or 24 h. HK2 mRNA and protein levels were detected via real-time PCR (a) and western blot (b). (c, d) Control and shYAP MCF7 cells were cultured with control medium or the indicated macrophage CM for 12 h. HK2 mRNA and protein levels were determined by real-time PCR (c) and western blot (d). (e) Control and shTEAD MCF7 cells were cultured in control medium or macrophage CM. The protein level of HK2 was determined by western blot. (f) MCF7 cells were transfected with siRNA for TNFR1 and then cultured in macrophage CM or treated with 10 μM TNFα. The cells were lysed for the detection of HK2 expression. (g, h) MCF7 cells stably expressing shRNA targeting HK2 were cultured in macrophage CM (g) or treated with 10 μM TNFα (h) in transwells for 24 h. Cell migration were analyzed by transwell assay. (i) Clinical specimens of breast cancer were immunostained with antibodies against CD68, YAP and HK2, the typical specimens of the same region were exhibited, the classification and the correlation between two proteins were performed. The error bars represent the means±s.d. (**P<0.01; ***P<0.001; n=3).