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. 2015 Jul 21;5:12278. doi: 10.1038/srep12278

Figure 2. TRPV1 tetrameric concatemer self-assembles into a functional receptor channel.

Figure 2

(a) Schematic illustration of the different tandem constructs used to study the assembly of rTRPV1 concatemers. Black squares represent wild type rTRPV1 subunit (wt) and red squares represent dominate-negative subunits (DN) mutated in positions NML676FAP. (b) Bar diagram represents the average (±SEM) amplitudes of a whole-cell, capsaicin-evoked (1 μM) currents from Xenopus laevis oocytes expressing the different tandem constructs either alone (black bars), or together with a DN construct (red bars; DN). Holding potential −60 mV. Each bar represents readings from 6–15 oocytes. The statistical significance between currents evoked with or without DN was determined using unpaired Student’s t tests, where **represents P ≤ 0.01, ***represents P ≤ 0.001. Note that no statistical significant difference in evoked currents was found between cells expressing 4wt with or without DN. (c) Western-blot analysis of HEK293T cells transiently expressing a single subunit (wt) or a tetrameric tandem (4wt) constructs, using an anti-rat TRPV1 antibody (α rTRPV1). Note that, while the expression of a single subunit wt construct yielded a band corresponded to ~90 kDa, the only specific band observed in cells expressing the 4wt construct corresponded to a ~400 kDa size (indicated with arrows).