Figure 4.

p53-R273H upregulates NRP2 to promote cell migration. (a) H1299 cells stably expressing mutant p53-R273H were subjected to RNA-seq based digital gene expression (DGE) analyses. Differentially expressed genes were identified by a P-value ⩽ 0.01 and an expression change of twofold or more between the two samples. (b,c) H1299 cells stably expressing p53-R273H were infected with recombinant lentivirus encoding DLX2. Cells were subjected to Q-PCR analysis (b) for NRP2 expression, and to western blot analyses (c) for NRP2, DLX2 and p53 expression. Results are presented as means and S.E. from three independent experiments in triplicates. ** indicated P<0.01. (d–g) Stable H1299 cells expressing p53-R273H were infected with recombinant lentivirus encoding a shRNA specific for NRP2 (shNRP2-1 or shNRP2-2) or a shGFP control were subjected to western blot analyses, as indicated (d); or to Q-PCR experiments to measure expression of NRP2, E-cadherin or Snail, as shown (e); or to transwell migration (f) or invasion (g) assays. Results are presented as means and S.E. from three independent experiments in triplicates. Scale bar =100 μm. ** indicated P<0.01