Journal: eLife

Article Title: Ancestral acetylcholine receptor β-subunit forms homopentamers that prime before opening spontaneously

doi: 10.7554/eLife.76504

Figure Lengend Snippet: Representative single-channel activity of β Anc homomers in the presence of increasing concentrations of ( A ) acetylcholine and ( B ) QX-222. Openings are upward deflections. Recordings were obtained with an applied voltage of –120 mV. Data were filtered to 10 kHz (scale bars = 25 ms, 10 pA; applies to (A) and (B)). The sequence of dwells from each dataset, encompassing the full concentration range of the blocker, was globally fit to the same three-state scheme used for β Anc , where an additional fourth state corresponding to the open/blocked channel was added . Global kinetic fits were performed on three individual recordings for each concentration of blocker, from at least two separate transfections, corresponding to 15 total patches for each global fit. Note that the recordings in the absence of blocker are the same for each dataset. Rate constants are overlaid on the scheme below each dataset, with error estimates presented in . Figure 4—source data 1. Source data for . Detected single-channel event durations of spontaneously opening β Anc homomers in the absence (agonist-free) and presence of increasing concentrations of acetylcholine (ACh; 10 µM, 30 µM, 60 µM, and 100 µM) or 2-[(2,6-dimethylphenyl)amino]- N , N , N -trimethyl-2-oxoethaniminium chloride (QX-222; QX; 10 µM, 30 µM, 60 µM, and 100µM). Compressed file includes 27 TAC 4.3.3 event files (*.evt format) of the single-channel detections for the three recordings for each condition (fileA, fileB, and fileC in each case) in the presented kinetic analysis, as well as the associated R scripts (*.txt format) for defining and sorting bursts. Figure 4—source data 2. Unrasterized version of .

Article Snippet: Chemical compound, drug , QX-222 , Tocris , 1043/10 , Purity:>98%.

Techniques: Activity Assay, Sequencing, Concentration Assay, Transfection