Figure 3
From: Live imaging-based assay for visualising species-specific interactions in gamete adhesion molecules
The accumulation and translocation of IZUMO1 and JUNO depended on their amino acid sequences. (A) LIAM assay with mouse IZUMO1/SPACA6 and JUNO. Mixing of IZUMO1-expressing cells (green) and JUNO-expressing cells (magenta) induced IZUMO1 accumulation (top panels) or simultaneous accumulation of IZUMO1 and JUNO (middle panels) (see Video 4). Arrowheads indicate accumulation of IZUMO1 or JUNO. Point mutations of amino acids critical for binding, i.e., W148A IZUMO1 (bottom left) and W62A JUNO (bottom middle), impaired accumulation. The sperm membrane protein SPACA6 (bottom right) did not induce accumulation of JUNO. Bars: 10 µm. (B) IZUMO1/SPACA6 and JUNO accumulation rates. (C) Translocation of IZUMO1 to an adjacent cell. Note that translocated signals (enclosed by dashed lines) continued to act as florescence foci. Bars: 10 µm. (D) IZUMO1/SPACA6 and JUNO translocation rates.
