FIGURE 8.

Analysis of the effect of B-Raf expression on NHE1 protein levels and targeting. A, surface localization of HeLa and HEK cells expressing NHE1 protein in the presence or absence of additional NHE1 protein. Cells were transfected with NHE1 in the presence or absence of B-Raf (or empty vector). Surface localization was determined as described under “Experimental Procedures.” Equal amounts of total cell lysate (T) and unbound intracellular lysate (U) were examined by Western blotting with anti-HA antibody to identify NHE1 protein. Ct and B-Raf refer to control cells or cells expressing B-Raf. The percent of the total NHE1 protein found on the plasma membrane is indicated. Results are mean ± S.E. n = at least four determinations. There was no significant difference between control and B-Raf-transfected cells. B, summary of Western blot analysis of NHE1 expression in control HeLa or HEK cells or in cells transfected with B-Raf as in A. Expression levels of experimentals were compared with that of controls. There was no significant difference between controls (Ct) and cells transfected with B-Raf.