Fig. 5. Expression of E-cadherin and N-cadherin in growth factor-stimulated thyrocytes. (A) Immunofluorescent staining of filter-cultured cells treated with TGF-ß1 (10 ng/ml) and EGF (10 ng/ml) for 24 hours. (Upper panel) E-cadherin (E-cad); (lower panel) N-cadherin (N-cad). Specimens were counterstained with DAPI (red) to visualise the cells' nuclei. N-cad was immunolocalised with clone CH-19, which is a pan-cadherin antibody raised against a sequence of the highly conserved cytoplasmic domain of N-cadherin. CH-19 gives results identical to that of the 3B9 monoclonal N-cadherin antibody in western blot analysis, but was used for immunofluorescence due to superior staining intensity. Bar, 15 µm. (B) Western blot analysis of E-cadherin and N-cadherin (using clone 3B9) after stimulation with TGF-ß1 (T; 10 ng/ml) and/or EGF (E; 10 ng/ml) for the indicated times. ß-actin indicates equivalent protein loading. (C) Appearance of distinct small size bands with E- or N-cadherin immunoreactivity after co-stimulation for 48 hours.

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