Binding assay between ZP-free eggs and HEK293T cells overexpressing Dcst1/2.
A Detection of DCST1/2 and IZUMO1. FLAG-tagged DCST1, HA-tagged DCST2, and 1D4-tagged IZUMO1 were detected in HEK293T cells overexpressing Dcst1-3xFLAG, Dcst2-3xHA, and Izumo1-1D4. B, C Observation of ZP-free eggs incubated with HEK293T cells overexpressing Dcst1/2 and Izumo1. The HEK293T cells overexpressing Dcst1 or Dcst2 did not attach to the oocyte membrane. Even when the HEK cells overexpressing Dcst1/2 were used for the assay, these cells failed to bind to ZP-free eggs. The HEK293T cells overexpressing Dcst1/2 and Izumo1 could bind to the oocyte membrane but could not fuse with an egg. n.s.: not significant (p = 0.28). The scale bars are 100 μm. All values are shown as the mean ± SD.
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