Identification of N-terminally deleted AFX protein, α(198-505).

<p>(A) Three start codons (M1, M2, and M3) are present in exon 1b and 2. M3 is the methionine codon located at the end of forkhead domain, which contains a consensus Kozak sequence for initiation of translation. (B) Western blot analysis. We detected an additional band (arrow) using anti-HA antibody from the lysate of 293-FT cells transfected with AFXα-HA (C-terminal tagged), suggesting a presence of N-terminally deleted AFX protein. (C) Immunoprecipitation analysis. To identify truncated protein, three N-terminally deleted AFX constructs tagged with C-terminal-HA epitope {α(131-505)HA, α(178-505)HA, and α(198-505)HA} were transfected. Western blotting with anti-AFX antibody (C-terminal specific) after immunoprecipitation with anti-HA antibody of transfected cell lysates revealed α(198-505) protein (arrow).