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. 2010 Dec 3;5(12):e14229. doi: 10.1371/journal.pone.0014229

Figure 5. PPARG silencing increases proliferation and migration/invasiveness of CRC cells.

Figure 5

(A) MTT assays on HCT116 and HT29 cells were carried out at different time-points *P = 0.004; **P = 0.001. (B) A wound-healing migration assay was carried out comparing and measuring the “wound area” at 24 and 48 hs *P = 0.045; **P = 0.0051; ***P = 0.0001. (C) Transwell migration assay was performed counting the run-through cells in 10 microscopic fields *P = 0.024; **P<0.01. The symbols represent the mean values of three independent experiments (mean ± SD). (D) Specific PPARG- or scrambled-shRNAs were stably transfected into HT29 cells to generate the shPPARG or control clones, respectively; the extent of PPARγ knock-down was documented by Western blot and referred to β-actin. (E) shPPARG cells showed higher proliferation than control clones and parental cells *P = 0.022; **P = 0.012. (F) The wound-healing migration and transwell migration assays were performed on the HT29 parental, the shPPARG and the control clone, respectively. The measurements were done as above. In both cases, cells were fixed after 48 hs and stained with hematoxylin & eosin or crystal violet, respectively. Magnification: 100×. Quantification of the wound-area after 24 and 48 hs is reported in the histogram where the control was set at 100%, *P = 0.016, **P = 0.001. Bars represent mean values ± SD of three independent experiments. (G) HCT116 cells were stably transfected with an empty expression vector or a vector carrying the PPARγ cDNA to generate control or the HCT116-PPARγ clones, respectively. Western blot analysis of the transfected PPARγ and activated target E-cadherin referred to β-actin. (H) The HCT116-PPARγ cells showed lower proliferation than control clones and parental cells in the presence of TZD *P = 0.012. (I) The wound-healing migration assay in HCT116 parental, the HCT116-PPARγ and the control clone. Cells were fixed after 48 hs and stained with hematoxylin & eosin. Magnification: 100×. The histogram shows quantification of the wound-area, measured as above, with the control set at 100%. Bars represent mean values ± SD of three independent experiments *P<0.05; **P<0.01.