Amino acid sequence comparisons of the three structural features of the gerbil, lizard and frog prestins.

<p>Id = identities; Pos = Positives.</p

Heterogenic expression and NLC measured from prestin orthologs on transfected HEK cells.

<p><i>(A)</i> Examples of confocal microscopy images of HEK cells transfected by lPres in the attached (left panel) and detached conditions (middle panel), and by fPres (right panel). Bar: 10 µm. <i>(B)</i> Means (heavy lines) and standard deviations (light color lines) of NLC obtained from HEK cells transfected by lPres and fPres, respectively. The mean capacitance-voltage responses were fitted with Boltzmann function. Linear capacitance (C_lin) was used to normalize NLC.</p

Examples of motile responses measured from HEK cells transfected by gPres, lPres, fPres, and EGFP vector, respectively.

<p>The cell was approximately 50% inserted into the microchamber and length change of the extruded segment was measured by a photodiode-based displacement-measurement system. The electrical stimulus (bottom panel) was a 100-Hz sinusoidal voltage burst with duration of 100 ms. No time registered motile response was seen from lPres-, fPres- and EGFP-transfected HEK cells. Motility responses measured were observed from gPres-transfected HEK cells. The responses were the results of 200 averages.</p

Alignment of consensus amino acid sequences of SLC26A5 of gerbil (Meriones unguiculatus, Mungu), frog (Xenopus tropicalis, Xtrop) and anole lizard (Anolis carolinensis, Acaro).

<p>Color of each residue represents identity at the residue among three different species. Blue: Full identity at a residue; Black: Partial identity (2/3 sequences) at a residue; Red: complete disparity at a residue. Gaps in the aligned sequences are indicated by the dashed line. The red box marks the motif that is remarkably conserved among mammalian species but highly variable among non-mammalian orthologs. This area may reflect a structural adaptation that facilitates gain of motor function in mammalian prestin.</p

Transport activity measured using radioisotope technique with [¹⁴C] formate as the substrate.

<p>Human pendrin was used as a positive control and EGFP plasmid was used as a negative control. Inhibition of formate uptakes by DIDS was also presented. All the data were acquired from 3 wells in each plate and repeated 3 times.</p

Voltage-dependent NLC of prestin and its orthologs.

<p>(<i>A</i>) Means of NLC responses measured from gPres (n = 11), pPres (n = 9), cPres (n = 12), lPres (n = 10) and fPres (n = 8). The mean capacitance-voltage responses were fitted with Boltzmann function. (<i>B</i>) Four parameters obtained from curving fitting using Boltzmann function.</p