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. 2023 Jan 16;14:235. doi: 10.1038/s41467-022-35705-4

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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. 3 — a qPCR analysis of islet amino acid sensing GPCRs (gprc6a, tas1r3, casr) (data represent the means ± SD, n = 4 for each group). b Quantification of α cell number in 5 dpf larvae of control, gcgr^DKO, and gcgr^DKO with knockdown of amino acid sensing GPCRs (gprc6a, tas1r3, casr) (Data represent means ± SD, n = 12, 12, 5, 8, 16 for each group). c. Diagram of casr CRISPR mutagenesis including target location, target sequence (underlined), PAM (red) and characterized mutations. Lower case letters indicate insertion. Hyphens represents deletion. d. Representative images of α cells in control, gcgr^DKO, cas+/-;gcgr^DKO and casr−/−;gcgr^DKO larvae at 5 dpf (scale bar, 10 μm). e Quantification of α cell numbers in control, gcgr^DKO, cas+/-;gcgr^DKO and casr−/−;gcgr^DKO larvae at 5 dpf (data represent the means ± SD, n = 12 for each group). f Free amino acid levels of control, casr−/−, gcgr^DKO, and casr−/−;gcgr^DKO larvae at 5 dpf, normalized by total protein concentration (data represent the means ± SD, n = 3/4, 40 fish/replicate, for each group). g Representative images of α cells in control and gcgr^DKO larvae treated with 1 μM NPS 2143 or vehicle for 48 h at 5 dpf (scale bar, 10 μm). h Quantification of α cell numbers in control and gcgr^DKO larvae treated with 1 μM NPS 2143 or vehicle for 48 h at 5 dpf (data represent the means ± SD, n = 12 for each group). i. Representative images of EdU staining in control and gcgr^DKO larvae treated with 1 μM NPS 2143 or vehicle for 48 h at 5 dpf. EdU (red)-positive α cells are indicated by arrows (scale bar, 10 μm). j. Quantification of EdU positive α cell numbers in control and gcgr^DKO larvae treated with 1 μM NPS 2143 or vehicle for 48 h at 5 dpf (data represent the means ± SD, n = 12 for each group). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, NS means no significant difference (two-tailed unpaired t test was used for Fig. 2a. One-way ANOVA and Tukey’s multiple comparisons test were used for Fig. 2b, e, f, h and j; the quantifications represent individual islet sections). Source data are provided as a Source Data file.