Extended Data Fig. 1: Expression of GFP-tagged NOMPC variants in Drosophila S2 cells.
From: NOMPC ion channel hinge forms a gating spring that initiates mechanosensation
a, Protein localization. GFP fluorescence reports cell surface localization for all three variants. n ≥ 100 cells/construct. b, Currents and respective channel numbers in excised outside-out membrane patches. Left: channel number \(N\) per patch deduced by fitting \({p}_{o,\ge 1}\left(P\right)\) (Fig. 1h) with \({Y\left(P\right)=1-\left(1-{p}_{o}\left(P\right)\right)}^{N}\). Middle: respective peak current amplitudes. Right: channel number deduced by dividing the peak current (middle) by the respective single channel current amplitude (Fig. 1d) (n = (n = 7 cells per protein variant except for n = 9 (NOMPC::GFP) and n = 6 (LH+LHCys-NOMPC::GFP)). Both methods yield similar channel numbers. c, Top: respective normalized amplitudes (\(I/{I}_{\max }\)) as function of the stimulus pressure (n = 7 cells per protein variant except for n = 9 (NOMPC::GFP) and n = 6 (AR + AR-NOMPC::GFP)). Solid lines: respective Boltzmann fits. Hatched lines: \({p}_{o}\left(P\right)\) deduced from Fig. 1f by fitting \({p}_{o,\ge 1}\left(P\right)\) with \({Y\left(P\right)=1-\left(1-{p}_{o}\left(P\right)\right)}^{N}\). Bottom: superimposed Boltzmann fits from the top panels and respective offset pressure corresponding to half-maximal current amplitude (shown for \({p}_{o}\left(P\right)\) in Fig. 1g). Compared to the deduced \({p}_{o}\left(P\right)\), the normalized current amplitudes increase less steeply with the stimulus pressure (top), yet, like \({p}_{o}\left(P\right)\), they superimpose for AR + AR-NOMPC::GFP and NOMPC::GFP and are shifted to approximately twice larger pressures for LH + LH-NOMPC::GFP and LH+LHCys-NOMPC::GFP.
