Figure 2.

ΔNp63α inhibits Hs-578T cell outgrowth in Matrigel and cell invasion. Hs-578T, H1299 and A549 cells were infected with a recombinant retrovirus expressing a vector control (C), WT murine ΔNp63α (WT) or a mutant ΔNp63α derivative (C306R or C526W), as shown, and selected by puromycin resistance. (a) Stable Hs-578T cells were seeded into 96-well plates for Matrigel outgrowth assay, and photographed after 7 days to assess morphology. Representative pictures from three independent experiments are shown. Scale bar=400 μm. (b) Stable Hs-578T, H1299 and A549 cells were subjected to transwell assays for cell invasion. Twenty-four hours after plating, invading cells were fixed and stained with crystal violet, and quantitated as described in the Materials and Methods section. Results are presented as means and S.E. from three independent experiments. ***P<0.001. (c) Whole-cell lysates from stable cells were subjected to western blotting, as indicated. (d) Stable cells were subjected to cell proliferation analysis by counting the number of cells every 24 h with a hemacytometer, as described in the Materials and Methods section. Results are presented as means and S.E. from three independent experiments. GAPDH, glyceraldehyde 3-phosphate dehydrogenase; PARP, poly (ADP-ribose) polymerase