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. 2023 Apr 3;18(6):667–676. doi: 10.1038/s41565-023-01359-6

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a, In vivo retinal fundus image showing MscL–tdTomato expression. b,c, Confocal stack projections across the RGC layer of a flat-mounted retina. d, Density of RBPMS-positive, MscL-positive and double-labelled cells (n = 5 WT MscL and G22S MscL retinas; *p = 0.0140, for RBPMS(+); *p = 0.0465 for RBPMS(+)/MscL(+), unpaired two-tailed t-test). e, Schematic of the experimental setup with an image of the retina on MEA electrodes. f, Representative peristimulus time histograms (PSTHs) for US or visual stimuli in MscL-transfected or NT RGCs (US stimuli, 15 MHz at 1.27 MPa). g, RGC response latencies to a 15 MHz US stimulus for MscL (n = 300 cells, 9 retinas) and NT retinas (n = 41 cells, 4 retinas). Dotted line, 45 ms latency threshold. h, Numbers of cells per retina responding to 15 MHz US stimuli (0.98–1.27 MPa) for MscL (n = 9 retinas) and NT (n = 4 retinas) with SL (<45 ms) or LL (>45 ms). *p = 0.0002, unpaired two-tailed t-test. i, Mean numbers of SL-responding RGCs per retina following stimulation with US stimuli of increasing pressures for MscL (n = 9) and NT (n = 4) retinas. ***p = 0.00008, ***p = 0.0010, ***p = 0.0008, multiple unpaired two-tailed t-tests. j, Maximum firing rates and response durations (SL and LL RGCs from MscL retinas in response to US stimuli of increasing pressures (0.20–1.27 MPa)) (n = 9 retinas, **p = 0.0017, *p = 0.0418, unpaired two-tailed t-test). k, Percentages of SL RGC cells (normalized against the maximum number of responsive cells in each experiment) responding to US stimuli for WT MscL (n = 3 retinas) and G22S MscL (n = 6 retinas) retinas. **p = 0.0065, **p = 0.0083, multiple unpaired two-tailed t-tests. l, Ratios of RGCs responding to US stimulation with SL or LL for MscL and NT retinas (n = 9 retinas for MscL and 4 retinas for NT), following the application of a cocktail of synaptic blockers (CNQX-CPP-LAP4, n = 3 retinas for both MscL and NT) and for P23H retinas with and without MscL expression (for both, n = 3 retinas). *Conditions with no US-elicited cell responses. Data are presented as mean values ± standard error of the mean (s.e.m.). Scale bars, 100 μm (b), 20 μm (c), 200 μm (e).

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