Journal: medRxiv

Article Title: Semaglutide alters the human embryo–endometrium interface

doi: 10.64898/2026.03.03.26347354

Figure Lengend Snippet: a–d , Representative images of immunohistochemical staining of GLP-1R (yellow) in endometrial tissue from healthy fertile women during the proliferative (P, N=5) ( a ), early secretory (ES, N=5) ( b ), mid-secretory (MS, N=5) ( c ), and late secretory (LS, N=5) ( d ) phases of the menstrual cycle. Scale bar, 100 µm and magnification 50 µm. e , Quantification of the GLP-1R positive glandular epithelial area fractions (%) for the P, ES, MS and LS phases of the endometrial tissue. Each violin represents distribution, median, and range of the feature for each group. Asterisks indicate statistically significant differences between the groups. Statistical significance was determined using a Kruskal-Wallis test followed by Dunn’s multiple comparisons test (*p < 0.05, ***p < 0.001, ****p < 0.0001). f, Representative immunofluorescence image of EEOs stained for GLP-1R (yellow) and Ki-67 (pink). Scale bar, 50 µm. g , Normalized ΔFRET% in endometrial epithelial cells treated with forskolin (Fsk; positive control) or semaglutide (SE) (0.039, 1.25, or 5□μM) in the absence or presence of estrogen and progesterone (EP). Error bars represent the Standard Error of the Mean (SEM) calculated from two independent biological replicates, with each experiment performed in technical triplicates. Statistical significance was determined using a one-way ANOVA followed by Dunnett’s multiple comparisons test (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001). h , Median expression obtained from 4 biological replicates of EEOs for endometrial receptivity genes LIF, PAEP, SPP1, GBP2, AQP3, DYNLT3, ANXA2 and YARS2 in EEOs untreated with SE (EEO), and treated with SE (EEO-SE 0.039 μM, 1.25 μM, or 5□μM) in absence of estrogen, progesterone and cAMP (EPC). Alongside expression was assessed in EPC stimulated EEOs without SE exposure (EEO-EPC) and with SE exposure (EEO-EPC-SE 0.039 μM, 1.25 μM, or 5□μM) (log□-transformed normalized counts). i , Heatmap of genes associated with metabolic pathways in EPC stimulated EEOs following SE treatment at 0.039 μM, 1.25 μM, or 5□μM (log□-transformed normalized counts). j–n , Oxygen consumption rate (OCR) measured by Seahorse XF Mito Stress Test in three biological replicates of EEOs treated with SE at 0.039 μM, 1.25 μM, or 5□μM, and SE untreated EEOs as control. Data represent mean ± SEM of three biological replicates, each measured in technical triplicates, with values normalized to DNA content. j , Pooled and normalized OCR profiles of EEOs treated with SE. Sequential injections of oligomycin, FCCP, and rotenone/antimycin A were used to assess mitochondrial function parameters. k–n , Quantification of OCR-derived parameters: k , basal respiration; l , maximal respiration; m , ATP production and n , proton leak. Error bars represent the SEM. Statistical analysis was performed using a one-way ANOVA followed by Dunnett’s multiple comparisons test (*p < 0.05, **p < 0.01, ****p < 0.0001).

Article Snippet: The dilutions of SE or positive control forskolin (Tocris Bioscience) on transparent 96-well clear-bottom plates (Thermo Fisher Scientific) were made into PBS containing 7.5 mol/L BSA (Sigma-Aldrich) to reduce non-specific binding.

Techniques: Immunohistochemical staining, Staining, Immunofluorescence, Positive Control, Expressing, Transformation Assay, Control, Derivative Assay

Journal: medRxiv

Article Title: Semaglutide alters the human embryo–endometrium interface

doi: 10.64898/2026.03.03.26347354

Figure Lengend Snippet: a , Normalized change in Förster resonance energy transfer efficiency (ΔFRET%) reporting intracellular cAMP levels in ESCs treated with forskolin (Fsk) or SE (0.039 μM, 1.25 μM, or 5□μM), with or without estrogen and progesterone (EP/ NO EP). b , Relative cellular metabolic activity of SE-treated (0.039 μM, 1.25 μM, or 5□μM) and untreated ESCs assessed by resazurin assay. c , Prolactin (PRL) secretion measured by ELISA after 9 days of EPC stimulation in SE-treated (EPC 0.039 μM, 1.25 μM, or 5□μM SE) and SE-untreated (ESC-EPC) ESCs. Error bars represent the Standard Error of the Mean (SEM) calculated from three independent biological replicates, with each experiment performed in technical duplicates. Statistical significance was determined using a one-way ANOVA followed by Dunnett’s multiple comparisons test (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001). d , Insulin-like Growth Factor Binding Protein 1 secretion (IGFBP-1) secretion measured by ELISA after 9 days of EPC stimulation in SE-treated (EPC 0.039 μM, 1.25 μM, or 5□μM SE) and SE-untreated (ESC-EPC) ESCs. Error bars represent the SEM calculated from four independent biological replicates, with each experiment performed in technical duplicates. Statistical significance was determined using a one-way ANOVA followed by Dunnett’s multiple comparisons test (***p < 0.001, ****p < 0.0001). e , Heatmap of G2/M phase-related genes in ESCs treated with EPC and SE (EPC-SE-0.039 μM, 1.25 μM, or 5□μM) (log - transformed normalized counts). f , Heatmap of genes associated with the unfolded protein response, N-glycan biosynthesis, and ER-associated degradation (ERAD) pathways in ESCs treated with EPC and SE (EPC-SE-0.039 μM, 1.25 μM, or 5□μM) (log□-transformed normalized counts).

Article Snippet: The dilutions of SE or positive control forskolin (Tocris Bioscience) on transparent 96-well clear-bottom plates (Thermo Fisher Scientific) were made into PBS containing 7.5 mol/L BSA (Sigma-Aldrich) to reduce non-specific binding.

Techniques: Förster Resonance Energy Transfer, Activity Assay, Resazurin Assay, Enzyme-linked Immunosorbent Assay, Binding Assay, Transformation Assay, Glycoproteomics