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Fig. 4 Figure 4. LITAF partially abolishes NEDD4-2–dependent down-regulation of INa in HEK cells. A, co-immunoprecipitation of cell lysates from HEK cells transfected with plasmids for NEDD4-2–FLAG and LITAF-HA or just NEDD4-2–FLAG for 48h (n = 3). Top panel, the top part shows an interaction between immunoprecipitated NEDD4-2–FLAG and co-precipitated LITAF-HA (the asterisk indicates the light chain of the IP capture antibody), whereas the bottom part displays input levels of NEDD4-2–FLAG and LITAF-HA. Bottom panel, respective changes in input NEDD4-2 expression, normalized to tubulin. All values are means ± S.E. (n = 3). B, current–voltage relationships of INa peak currents for baseline conditions from cells expressing. GFP (control), NEDD4-2, or NEDD4-2 and LITAF (means ± S.E.). Two-way analysis of variance for repeated measures revealed significant differences in INa between all groups: GFP versus NEDD4-2: F = 80.05, p < 0.0001. NEDD4-2 versus NEDD4-2 + LITAF: F = 11.42, p = 0.0008. GFP versus NEDD4-2 + LITAF: F = 15.28, p = 0.0001.