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Figure 1 | Molecular Brain

Figure 1

From: Depletion of 14-3-3γ reduces the surface expression of Transient Receptor Potential Melastatin 4b (TRPM4b) Channels and attenuates TRPM4b-mediated glutamate-induced neuronal cell death

Figure 1

TRPM4b associates with 14-3-3γ in heterologous systems. (A) Diagram of the full length TRPM4b, a splice variant TRPM4a, and a short TRPM4b N-terminal fragment (TRPM4b-N174) used in this study. (B) Identification of 14-3-3γ as a direct binding partner of TRPM4b in the yeast two-hybrid system. (C) Co-IP experiment. GFP-TRPM4b (4b) or GFP-TRPM4b-N174 (N174) was transfected with or without FLAG-14-3-3γ in HEK293T cells and cell lysates were then immunoprecipitated using anti-FLAG antibody. Immunoprecipitates were examined by Western blotting using anti-GFP antibody (upper panel). Input represented 5% of cell lysates used in the Co-IP experiment (lower panel). (D) BiFC experiment with VN-14-3-3γ and VC-TRPM4b, where N- and C-terminal halves of Venus fluorescent protein were fused to 14-3-3γ and TRPM4b, respectively. Intense fluorescent signals were detected by the yellow color (arrow). Scale bar, 10 μm. (E) 14-3-3γ specifically binds to GFP-TRPM4b-N174 among seven 14-3-3 isoforms.

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Molecular Brain

ISSN: 1756-6606