FIG. 7.

(A) Coexpression of EphA8 and Odin in Neuro2a cells. One hundred micrograms of whole-cell extract (WCL) from PC12 or Neuro2a cells was resolved by SDS-PAGE and then subjected to Western blot analysis using the indicated antibodies. Note that Odin was detected as a smaller protein in PC12 cells than in Neuro2a cells. This may be because PC12 cells predominantly express a 1,125-amino-acid rat Odin protein, which is 25 amino acids shorter than mouse Odin in Neuro2a cells. (B) Coimmunoprecipitation of Odin and EphA8 in Neuro2a cells. Neuro2a cells stably expressing the WT EphA8 receptor were generated as described previously. Total cell lysates (500 μg) were immunoprecipitated with anti-Odin antibody and then immunoblotted with the anti-EphA8 antibody (top panel). The same blot was stripped and reprobed with anti-Odin antibody (middle panel). A sample (0.5%) of each total cell lysate was analyzed directly by Western blotting using anti-EphA8 antibody (bottom panel). (C) Odin colocalizes with EphA8 in the neurite tips of Neuro2a cells. Neuro2a cells stably expressing the EphA8 receptor were stained with either anti-EphA8 or anti-Odin antibody followed by Alex Fluor 594-anti-rabbit secondary antibody (top panels). Neuro2a cells transiently transfected with EphA8-EGFP were stained with anti-Odin antibody followed by Alex Fluor 594-anti-rabbit secondary antibody (bottom panels). WB, Western blot.