Figure 6.

Binding of EhSNX1 with the retromer complex. (a) Verification of the expression of EhSNX1‐HA by immunoblot analysis using anti‐HA antibody. Approximately 20 μg of total lysates from these strains were loaded. Cysteine synthase 1 was detected by anti‐CS1 antiserum as a loading control. (b) Immunoprecipitation of EhVps26 and EhSNX1‐HA followed by immunoblot analysis. After 30 min co‐culture with EhSNX1‐HA, HA‐EhSNX2, or mock transformant cells with CHO cells, the amoebae were collected to produce lysates, which were subjected to immunoprecipitation by anti‐Vps26 antiserum. The immunoprecipitated (“Bound”), unbound supernatant (“Unbound”), and non‐specific (protein G‐Sepharose control beads that were preincubated with lysates, washed, and boiled with SDS loading buffer) fractions were analysed with SDS‐PAGE and immunoblot analyses using anti‐HA antibody, anti‐Vps26, and anti‐Vps35 antisera