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. 2016 Apr 12;291(23):12322–12335. doi: 10.1074/jbc.M116.721191

FIGURE 5.

FIGURE 5.

R288A mutation in HJV and R281A mutation in Hjv disrupt the interaction with neogenin, but not with HFE or TfR2. A, co-immunoprecipitation of WT, R332A, and R288A-HJV with HFE and TfR2. HeLa-TfR2/fHFE cells were transfected by pcDNA3 (C), pcDNA3-HJV, R332A-HJV, and R288A-HJV, immunoprecipitated (IP) by using anti-HJV antibody, and immunodetected for TfR2, HJV, and fHFE. The entire images are illustrated in supplemental Fig. S1C. B, co-immunoprecipitation of WT, R281A, and R325A-fHjv with neogenin. HEK293 cells were co-transfected by pcDNA3-neogenin with pCMV9-fHjv, R281A-fHjv, or R325A-fHjv, immunoprecipitated by using anti-FLAG M2 beads, and immunodetected for neogenin (Neo) and fHjv. HEK293 cells transfected with pCMV9 alone pCMV9 (C) were included as a control. The entire images are illustrated in supplemental Fig. S1A. C, co-immunoprecipitation of WT, R332A, and R288A-HJV with neogenin. HEK293-neogenin (HEK-Neo) cells were transfected by pcDNA3 (C), pcDNA3-HJV, R332A-HJV, and R288A-HJV, immunoprecipitated by using anti-HJV antibody, and immunodetected for neogenin (Neo) and HJV. HEK293 cells (HEK) were included as a control. All experiments were repeated at least three times with consistent results.