Fig 1: Validation of the method with a comparison between native clusters and those obtained with overexpression of a construct with the N2B signature at the N-terminal and GFP at the C-terminal.(A) Four clusters found in the dataset of force-distance (F-D) traces pulled from NG108-15 cells wild type. (B) The Bayesian identification of the clusters in (A) with the candidate membrane proteins. (C) Confocal images of NG108-15 cells overexpressing the construct N2b-Protein-GFP, as reported by the green fluorescence emitted by GFP. (D) From left to right, color density plots (blue indicating a colocalization among F-D traces of more than 80%) of the clusters superimposed to an F-D curve bearing the N2B signature (85 nm segment with a flat force below 10 pN) obtained in the sample overexpressing the candidate protein TMEM16F, TRPC1, TRPC5, and TRPC6, respectively. (E) Global histogram of Lc of the clusters in A and the clusters with the N2B signature from cells transfected with the corresponding construct. (F) Position of the most likely rupture regions of the proteins according to (E) superimposed to the cartoon of the cryo electron microscopy (cryo-EM) structure available in the PDB.