ZFIN Image: Motta et al., 2021, Fig. 4

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Figure Caption

Fig. 4

p.Leu100Pro results in a defective SPRED2 binding to neurofibromin

(A) Subcellular localization of transiently expressed V5-tagged GRD and Xpress-tagged WT or SPRED2^Leu100Pro (L100P) proteins in COS-1 cells after stimulation with EGF (30 ng/mL, 5 min) revealed by confocal microscopy analysis. Cells were stained using anti-Xpress IgG1 and anti-V5 IgG2a monoclonal antibodies and revealed by goat anti-mouse IgG1 Alexa Fluor 647 (pseudocolored in green) and goat anti-mouse IgG2a Alexa Fluor 594 (red) secondary antibodies, respectively. Nuclei are visualized by DAPI staining (blue). Scale bar, 10 μm.

(B) Co-immunoprecipitation assays. Lysates from HEK293T cells transiently transfected to express WT or variant Xpress-tagged SPRED2 protein were serum-starved, stimulated with EGF (30 ng/mL), and immunoprecipitated with an anti-Xpress antibody and assayed by western blotting using the indicated antibodies.

Acknowledgments

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