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. 2013 Feb 28;8(2):e57829. doi: 10.1371/journal.pone.0057829

Figure 2. Knockdown of Wt1 and Tbx18 in primary epicardial cells.

Figure 2

(A) Representative images of primary epicardial cells transfected with control siRNA (siControl) or siRNA directed against Tbx18 (siTbx18) or Wt1 (siWt1). Scale bar: 200 µm. (B) Immunostaining for ZO-1 or N-cadherin (green) and DAPI nuclear staining (blue) of primary epicardial cells transfected with siRNAs. Scale bars: 100 µm. (C) Percentage of cells categorized as “Enlarged” or “Cobblestone-like,” based on the cellular morphology of primary epicardial cells transfected with siRNAs. (D) Representative images of primary epicardial cells transfected with siRNAs at 0 and 14 hr after the scratch was made. Scale bar: 200 µm. (E) Quantification of migration distance, given as a ratio to the siControl (n = 4; *P<0.01 vs. siControl). (F) The relative mRNA expression of Tbx18, Wt1, Snail and Slug by real-time PCR analysis (n = 3; *P<0.05 vs. siControl). The results were normalized to Gapdh expression, and the relative expression level is given as a ratio to the siControl. (G) Western blot performed with antibodies against Tbx18, Wt1, adhesion molecules (E-cadherin and N-cadherin) and EMT regulators (Snail and Slug). β-actin and histone H3 were used as loading controls. The data are presented as the mean ± SD.