Figure 3. Effect of sodium selenate on Lm332 deposition by NHK cells.
(A) Immunoblotting analysis. NHK cells were inoculated in serum-free medium at a density of 4×105 cells per 35-mm dish, incubated overnight, and treated with (+) or without (−) 0.1 mM sodium selenate (Sigma) at 37°C for 24 h. After the incubation, the ECM and CM were prepared from each culture and analyzed for the laminin α3 chain by immunoblotting as described in Figure 1. (B) The ECMs from the control (none) and selenate-treated cultures (+selenate) were subjected to immunofluorescence staining with the anti-laminin α3 chain antibody BG5. (C) Phase-contrast micrographs of control and selenate-treated cultures. Other experimental conditions are described in “Materials and Methods”.