Skip to main content
Figure 2 | Molecular Brain

Figure 2

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 2

Interaction of TRPM4b with 14-3-3γ may require a conserved serine phosphorylation residue. (A) Schematic diagram depicting the membrane topology of hTRPM4b channels. Conserved phosphorylation residues (T19, T42, and S88) predicted for 14-3-3 binding were marked in the intracellular N-terminal domain (N174). These residues are highlighted in red. (B) GST-pulldown was performed on lysates of HEK293T cells co-expressing GST-14-3-3γ and GFP-TRPM4b-N174 or each mutant (S88A, T19A, and T42A). Cell lysates were immunoprecipitated using anti-GST antibody and immunoprecipitates were examined by Western blotting using anti-GFP or anti-GST antibody. Input represented 5% of cell lysates used. (C) Co-IP experiment was performed on lysates of HEK293T cells co-expressing FLAG-TRPM4b (WT) and its mutant (S88A). Cell lysates were immunoprecipitated using anti-FLAG antibody and immunoprecipitates were examined by Western blotting using anti-GFP or anti-FLAG antibody. Input represented 5% of cell lysates. (D) Sequence comparison of the region of conserved phosphorylation sites for 14-3-3 binding in hTRPM4b among human, rat, mouse and Xenopus. Only Ser88 was conserved over the species.

Back to article page

Molecular Brain

ISSN: 1756-6606