Fig. 3.

Interaction between IZUMO1 and GLIPR1L1 using reciprocal co-immunoprecipitation (IP). Acrosome-reacted mouse spermatozoa were subjected to immunoprecipitation using IZUMO1 or GLIPR1L1 antibodies as described in the “Methods” section. Membranes were probed with the target antibody, to confirm the efficacy of immunoprecipitation, before being stripped and re-probed with the alterative antibody to confirm the target protein interaction. Whole sperm lysate was included to confirm the identity of the co-precipitated proteins, as was the material recovered after washing the beads to confirm the specificity of the elution. This experiment was replicated three times and representative blots are depicted