Figure 4: ZNF304 mediated inside-out signalling.

(a) The anoikis rates of HeyA8 cells at 72 hours post transfection in suspension condition. HeyA8 cells were transfected with indicated siRNAs, and incubated in anoikis plates for 72 h; cells were analysed by flow cytometry following Annexin V-FITC and propidium iodide (PI) staining. Data are presented as mean±s.e.m. of n≥3 experimental groups. *P≥0.05, **P≥0.01,***P≥0.001, ****P≥0.0001 (Student’s t-test). (b) Immunoblotting analysis of HeyA8 cells in suspension conditions at 72 h after treatment with control siRNA or ZNF304 siRNA; followed by protein isolation and immunoblotting. Data are presented as mean±s.e.m. of n≥3 experimental groups. **P≥0.01,***P≥0.001, ****P≥0.0001 (Student’s t-test). Poly ADP ribose polymerase cleavage indicates enhanced anoikis in ZNF304 siRNA-treated and control siRNA-treated samples in suspension conditions. (c) The anoikis rates of HeyA8 and SKOV3IP1 cells in suspension after ZNF304 silencing and β1 integrin overexpression. Cells were analysed by flow cytometry following Annexin V-FITC and PI staining. Data are presented as mean±s.e.m. of n≥3 experimental groups. *P≥0.05, **P≥0.01,***P≥0.001 (Student’s t-test). NS, not significant.