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Fig. 3. Functionality of the CNTF-R and the LIF-R in MDCK cells. Parental MDCK (upper panel), stably transfected MDCK-CNTF-R (middle panel) or MDCK-gp130 cells (lower panel) were grown on Transwell filters for 5 days. The cells were apically stimulated with CNTF (25 ng/ml) or LIF (50 ng/ml) for 15-30 minutes at 37°C. Nuclear extracts were prepared and equal protein aliquots analysed by SDS-PAGE and western blotting. Phosphorylated STAT3 proteins were detected with an activation-specific STAT3 antibody (pY-STAT3), an HRP-conjugated secondary antibody and visualised using the ECL+plus system. Quantitation over basal levels (fold stimulation) was calculated in each set of experiment as indicated in the figure.





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