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. 2023 Dec 11;14:8207. doi: 10.1038/s41467-023-44040-1

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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. 10 — a Intracellular calcium levels were measured using Fluo-4 AM dye in HEK293T cells transiently transfected with control or overexpressing plasmids for CHRM3 and/or IL-31RA plasmids for 72 h and treated with 10 µM carbachol (n = 4). b Intracellular calcium levels were measured using Fluo-4 AM dye in HEK293T cells transiently transfected with control or overexpressing plasmid for IL-31RA for 72 h and treated with 50 µM bradykinin (n = 3). c, d, e, f Intracellular calcium levels were measured using Fluo-4 AM dye in hTERT cells transiently transfected with control or overexpressing plasmid for IL-31RA for 72 h and cells treated with 10 µM carbachol (n = 8), 10 µM bradykinin (n = 8), 100 µM serotonin (n = 4) or 10 µM ionomycin (n = 4). g hTERT cell line 72 (CHRM3^LOW) was transiently transfected with control or IL-31RA overexpression plasmids for 72 h and intracellular calcium levels were measured in cells treated with 10 µM carbachol (n = 5) or 10 µM ionomycin (n = 3) using Fluo 4 AM dye. Data are shown as means ± SEM. h Intracellular calcium levels were measured using Fluo-4 AM dye in hTERT cells transiently transfected with IL-31RA-specific or control siRNA for 72 h and treated with 10 µM carbachol (n = 8). i hTERT cells were transiently transfected with IL-31RA overexpressing plasmid for 48 h and treated with IL-31 (500 ng/mL) or media for another 24 h. Media or IL-31 treated cells were stimulated with 10 µM carbachol (n = 4) and intracellular calcium levels were measured using Fluo-4 AM dye. Data are shown as means ± SEM. j hTERT cells were transiently transfected with IL-31RA-specific or control siRNA for 72 h and treated with 10 µM carbachol (n = 3) for 10 min. Cell lysates were immunoblotted with antibodies against phospho-MLC, and β-actin. Data are shown as means $\pm$ SEM. Unpaired t test was used. k ASMC isolated from wildtype and IL-31RA^–/– mice were treated with 10 µM carbachol (n = 3) for 10 min and cell lysates were immunoblotted with antibodies against phospho-MLC, total-MLC and GAPDH. Data are shown as means $\pm$ SEM. Unpaired t test was used. At least two independent experiments produced similar results. Source data are provided as a Source Data file.