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. 2011 Oct 28;6(10):e26815. doi: 10.1371/journal.pone.0026815

Figure 7. A distinct role of ΔNp63α in HEK293 and H1299 cells.

Figure 7

A, ΔNp63α does not affect EGFR or 14-3-3σ protein levels in HEK293 or H1299 cells. Representative blot of three independent experiments is shown. B, ΔNp63α fails to upregulate EGFR promoter but modulates 14-3-3σ promoter in a luciferase reporter assay. Cells were transfected with wildtype EGFR or 14-3-3σ promoter luciferase construct as indicated, along with pCMVβ plasmid, with or without wildtype p53, ΔNp63α, ΔNp63αDBDmut, or TAp63α. At 48–72 h after transfection, the luciferase activity was determined. The transfection efficiency was standardized against β-galactosidase activity. Results are indicative of three independent experiments performed in duplicates. C, ΔNp63α increases 14-3-3σ mRNA transcript levels in H1299 cells. H1299 cells were transfected with ΔNp63α, ΔNp63αDBDmut or vector control. At 36 hours total RNA was isolated, reverse-transcribed and subjected to real-time PCR with probe specific for 14-3-3σ. Results were normalized to 18S levels. Data are the mean of two independent experiments done in duplicates in which similar results were obtained. D, ΔNp63α does not enhance anchorage-independent growth, migration or invasion in H1299 cells. H1299 cells were transfected with ΔNp63α or vector control. Soft agar, transwell migration and invasion assays were performed as described above. *, p<0.05 compared with control.