Extended Data Fig. 3: Data processing of OSCA1.2 in liposome.
From: Mechanical activation opens a lipid-lined pore in OSCA ion channels
a, Representative inside-out recordings of OSCA1.1 (blue, n = 6), OSCA1.2 (red, n = 6) and OSCA3.1 (olive, n = 6) expressed in Piezo1−/− HEK293T cells at −40 mV holding potential with vector only transfected cells as a negative control (black, n = 6). Inside-out patches were perfused with 250 µM of lysophosphatidylcholine 12:0 (LPC12:0) which elicited gating of OSCA1.1 and OSCA1.2 in the absence of applied pressure but failed to generate any current prior to patch rupture in OSCA3.1 or vector only transfected cells. Quantification of peak currents elicited by LPC12:0 from replicate cells are shown on the right side. The error bars represent the S.E.M. The p-values were determined using one way ANOVA with Dunnett’s multiple comparison. b, Image-processing workflow of OSCA1.2-azolectin-1:40-LPC with Relion and CryoSPARC. See Methods and Extended Data Table 1, 2 for details. c, Gold-standard FSC curves calculated between the independently refined half maps of OSCA1.2-azolectin-1:40-LPC in the expanded state indicate a resolution of 4.01 Å (FSC = 0.143) and in the contracted2 state indicate a resolution of 3.83 Å (FSC = 0.143). d, A representative cryo-EM image of OSCA1.2-liposome. Scale bar: 50 nm. e, Image-processing workflow of OSCA1.2-liposome with Relion and CryoSPARC. See Methods and Extended Data Table 1, 2 for details. f, Gold-standard FSC curves calculated between the independently refined half maps of OSCA1.2 in inside-in orientation indicate a resolution of 3.29 Å (FSC = 0.143) and in inside-out orientation indicate a resolution of 3.56 Å (FSC = 0.143). g, Cryo-EM map (upper panel, top view) and atomic model (lower panel, side view) of OSCA1.2 in inside-out and inside-in orientation. The membrane density is shown in transparent orange. h, Membrane blocks H3-H4 inwardly movement. The atomic model of OSCA1.2 closed subunit (grey) is aligned to OSCA1.2-inside-in open subunit (forest green). The membrane density is shown in transparent orange. i, TM6b moved 5 Å outward from closed state to open state.
