Figure 7.

rMMP-3 proteolytic domain-induced EMT in NRK52e cells is Slug dependent. A–I: Immunofluorescence images of NRK52e cells transfected with stealth negative control (ctr) siRNA (100 pmol) (A, D, G) or Slug siRNA oligonucleotides (50 pmol) (B, E, H) after treatment with rMMP-3 proteolytic domain (2 μg ml⁻¹) for 72 hours. Compared with untreated cells (C, F, I), EMT was induced in NRK52e cells transfected with stealth negative control siRNA after rMMP-3 proteolytic domain treatment, as demonstrated by loss of E-cadherin (A), de novo expression of α-SMA (D: Scale bar = 10 μm), and cytoplasmic and nuclear staining of β-catenin (G), but not in NRK52e cells transfected with Slug siRNA where E-cadherin (B) and β-catenin cytoplasm membrane staining (H) remained, and no α-SMA staining was observed (E). J and K: Western blot analysis of E-cadherin, α-SMA and Slug in lysates of NRK52e cells transfected with negative control (ctr) or Slug siRNA or no oligonucleotides after treatment with rMMP-3 proteolytic domain for 3 days. *P < 0.05.