Fig. 2. (A) Non-polarised expression of LIF-R in MDCK cells. HeLa, HepG2, Ba/F3-gp130AA, and Ba/F3-LIF-R cells or MDCK cells were extracted with lysis buffer and cell lysates immunoprecipitated with a monoclonal LIF-R specific antibody (7G7) or a LIF-R-specific rabbit antiserum, respectively. (B) MDCK and HeLa cells were grown on tissue culture dishes, lysed and incubated either with a LIF-R-specific rabbit antiserum [
-LIF-R (rb) +] or non-immune rabbit serum (rabbit serum +). (C) MDCK cells were grown on Transwell filters for 5 days. Sulfo-NHS-biotin was employed to selectively label the apical (ap) or the basolateral (bl) surfaces or cells were left untreated (ctr). The lysates were incubated with a LIF-R-specific rabbit antiserum. Immunoprecipitates (A-C) were analysed by SDS-PAGE and western blotting using either a polyclonal LIF-R-specific antibody followed by an appropriate HRP-conjugated secondary antibody (A,B) or HRP-conjugated streptavidin (C). Blots were developed using ECL+plus.