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. 2016 Oct 3;7:12939. doi: 10.1038/ncomms12939

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Copyright © 2016, The Author(s)

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(a) Measured force on fluorescently labelled magnetic nanoparticles (red, 75 nm diameter; n=15 nanoparticles per size). (b) Measured force on fluorescently labelled 75 nm diameter magnetic nanoparticles under varying current supply to electromagnet (red, 1 A and 5 A analyzed in next panel; n=15 nanoparticles per condition). (c) Inactivation and averages of individual experiments (top) and average differences (bottom) of BBS-2422 with ∼25% (1 A; 2.6±0.2 pN; n=8 cells, 4 transfections) and 100% (5 A; 9.9±0.7 pN; n=21 cells, 7 transfections) magnetic force with a −60 mmHg pressure step. (d) Percent inactivation and averages of individual experiments (top) and average differences (bottom) of construct BBS-2422 labelled with nanoparticles without magnetic field (n=8 cells each, 2 transfections) or unlabelled and with magnetic field with a −60 mm Hg pressure step (n=9, 2 transfections). (e) Schematic of antibody labelling on Piezo1-myc-2422 (pIRES-EGFP) and representative images of HEK293T cells expressing wild-type Piezo1 (pIRES-EGFP) or Piezo1-myc-2422 construct (green, GFP; red, anti-myc). Per cent inactivation and averages of individual experiments (top right) and average differences (bottom right) of myc construct labelled and unlabelled with anti-myc antibody (n=8 cells each, 2 transfections; NS, P>0.01, unpaired t-test). (f) Schematic of K_ir2.1-BBS construct labelled with nanoparticles co-expressed with wild-type Piezo1 (pIRES-EGFP) and representative images of surface labelled HEK293T cells (green, GFP; red, anti-streptavidin; grey, WGA). Inactivation and averages of individual experiments (top right) and average differences (bottom right) for co-expressed K_ir2.1-BBS and wild-type Piezo1 before and on magnetic force (n=10 cells, 2 transfections). Scale bar, 30 μm. (g) Inactivation and averages of individual experiments (top) and average differences (bottom) for BBS-2422 recorded first without magnetic field, second with magnetic field, and third again without magnetic field (n=9 cells, 5 transfections). (h) Representative single-channel recordings for BBS-2422 at resting tension with and without a magnetic field and quantification of single-channel conductance (n=9 cells, 4 transfections). Error bars are s.e.m. for all panels.