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. 2025 Jul 21;16:6687. doi: 10.1038/s41467-025-61852-5

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© The Author(s) 2025

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 9 — a Immunofluorescence of human pancreatic sections (IHC-IF) from pregnant women at different gestational ages and a non-pregnant control sample. The GLP-1 signal is observed in α cells, overlapping with glucagon. Both α cells surrounding pancreatic ducts and those within islets are shown. The grayscale images represent the individual channels for the GLP-1, glucagon and insulin. GLP-1 (green), glucagon (yellow), and insulin (magenta) are shown in the composite merged image. Scale bar = 200 µm. Representative images from four donors are shown; similar results were observed across the 14 biological replicates, each representing an independent human donor. b, c Images demonstrating the use of the imaging pipeline to quantify GLP-1 area and signal intensity. The labelling of GLP-1 labelled α cells by IHC-IF was followed by thresholding of the GLP-1 positive signal. An example of the thresholded region (cyan) identifying GLP-1 positive signal is shown for the (b) GLP-1 channel (green) and (c) composite merged image (GLP-1 [green], glucagon [yellow] and insulin [magenta]). d–h Quantitative comparisons of GLP-1 between pregnant and non-pregnant women for several quantified islet measures. The data includes comparisons for d) fractional area (measured area as a percentage of total tissue area) and (e) mean area (measured area divided by number of islets per tissue section). Additionally, comparisons of the proportions of GLP-1 positive area relative to (f) whole islets (measured area as a percentage of total whole islet area) and h) (g) relative to α cell area (measured area as a percentage of α cell area) are shown. h Signal intensity of GLP-1 detected in α cells of pregnant women compared to non-pregnant controls. Symbols in each figure correspond to individual donors as indicated in Supp. Data S1. The pregnant group (n = 7 biological replicates) was compared to the non-pregnant control group (n = 7 biological replicates). Each biological replicate represents an independent human donor. Data are presented as mean ± SEM. Normally distributed data were analysed using a two-sided unpaired Student’s t-test; non-parametric data were analysed using a two-sided Mann–Whitney test. Exact P values for each comparison are shown in the figure. Statistical significance was defined as P < 0.05. * GA gestational age, wks weeks.