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. 2013 Dec 27;8(12):e85624. doi: 10.1371/journal.pone.0085624

Figure 2. Overexpression of mutant p53-R261H disrupted tubular formation in 3-D culture.

Figure 2

A, Generation of MDCK cell lines in which siRNA-resistant mutant p53-R261H was stably overexpressed (clones 1 and 2). The protein levels of mutant p53-R261H and actin were measured by Western blotting. B, The level of wild-type p53 transcripts was determined by RT-PCR. C, Representative images of MDCK cells, MDCK cells with p53 knockdown, or MDCK cells with mutant p53-R261H in 2-D culture (200×). D, Representative images of MDCK cells with mutant p53-R261H in 3-D culture. Scale bar: 100 µM. E, Top panel: colony formation assay was performed with MDCK cells or MDCK cells with mutant p53-R261H. Bottom panel: the number of colonies was counted and presented as Mean ± SD from three separate experiments. F, Wound healing assay was performed with MDCK cells, MDCK cells with p53-KD, or MDCK cells with mutant p53-R261H. Top panel: cell migration was determined by visual assessment of cells migrating into the wound for 24 h using a phase-contrast microscopy. Bottom panel: the time required for wound closure was measured and presented as mean ± SD from three separate experiments.