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. 2012 May 14;32(10):1252–1265. doi: 10.1038/onc.2012.148

Figure 4.

Figure 4

Mutant p53 promotes MET-dependent invasion towards HGF. (a) Mutant p53 (273H) or control (EV) H1299 cells were analysed for invasion capacity in fibronectin-supplemented Matrigel using EGF or HGF as a chemo-attractant (left panel). Invasion was quantified as described in the Materials and methods section and values are means±s.e.m. of six replicates from each of three independent experiments (right panel). *indicates statistical significant changes (P<0.05) as determined by a T-test. (b) EI 175H H1299 cells were analysed for invasion capacity in fibronectin-supplemented Matrigel using HGF as a chemo-attractant after induction with increasing doses of ponA. Invasion was quantified as described in the Materials and methods section and values are means±s.e.m. of six replicates from each of three independent experiments (right panel). * indicates statistical significant changes (P<0.05) as determined by a T-test. (c) Mutant p53 (273H) or control (EV) H1299 cells were analysed for invasion after knockdown of MET using HGF as a chemo-attractant. * indicates statistical significant changes (P<0.05) as determined by a T-test. Knockdown of MET was verified by western blot (right panel), using GCN5 as a loading control. (d) Mutant p53 (273H) or control (EV) H1299 cells were analysed for invasion capacity in fibronectin-supplemented Matrigel with monoclonal antibodies against α5 integrin, using HGF as a chemo-attractant. * indicates statistical significant changes (P<0.05) as determined by a T-test. (e) Mutant p53 (273H) or control (EV) H1299 cells were transfected with RCP siRNA and analysed for invasion capacity in fibronectin-supplemented Matrigel using HGF as a chemo-attractant. Knockdown of RCP was verified by western blot using actin as loading control (right panel). * indicates statistical significant changes (P<0.05) as determined by a T-test. (f) H1299 EV cells were transfected with siRNA against ZO-1 or PAR3 and compared with 273H cells for invasion capacity in fibronectin-supplemented Matrigel using HGF as chemo-attractant. * indicates statistical significant changes (P<0.05) as determined by a T-test. (g) 3D reconstructive images of 273H H1299 cells invading towards EGF or HGF. (h) MDA MB231 cells were transfected with siRNA-targeting (mutant) p53 and analysed for invasion capacity into fibronectin-supplemented Matrigel using HGF as chemo-attractant (middle panel). Knockdown was verified by western blot using actin as loading control (left panel). Quantified invasion is shown in the right panel.