Figure 9.

Bufalin-induced changes in cellular distribution of Na⁺, K⁺-ATPase α subunits in NT2 cells. NT2 cells were grown on glass coverslips for 24 h. The DMEM-F12 was then replaced with medium with (D-F) or without (A-C) 20 nM bufalin for 4.5 h. The cells were fixed and stained with anti Na⁺, K⁺-ATPase α subunits monoclonal antibodies, and images were acquired by fluorescence microscopy (B and E) were obtained as described in MATERIALS AND METHODS. The merge of the phase contrast and fluorescence is depicted in C and F. Bar, 10 μm. Quantification of the distribution of the Na⁺, K⁺-ATPase α subunits was performed as described in the legend to Figure 5 and is shown in G. The total cellular Na⁺, K⁺-ATPase α subunits (H) were assessed by Western blot analysis and quantified by densitometry.