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Binding of mRSPO1-Fc to LGR4 and LGR5 by confocal immunofluorescence analysis. HEK293 cells stably expressing vector, Myc-LGR4, Myc-LGR5, or HA-LRP6 were incubated with mRSPO1-Fc at 4 °C (A), or at 37 °C (B). The cells were then costained with fluorescence-labeled anti-tag antibodies (Cy3–anti-Myc, Alexa594–anti-HA, both mouse IgG1 subtype) for receptor detection (red) and Alexa Fluor 488-labeled anti-IgG2a for mRSPO1-Fc detection (green). Nuclei were counterstained with ToPro-3 (blue).
