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Proteomic analysis of Ishikawa-sEVs. A Timeline of the experimental protocol for hormonal treatment of Ishikawa cells. B Concentration of sEVs (µg/mL; total protein amount of sEVs per volume of conditioned culture medium) in the two groups. Data are presented as mean ± SD. Statistical significance was assessed using the Student’s t -test (ns, not significant). C Dot plots showing the expression levels of proteins identified in Ishikawa-sEVs (915 proteins) ranked in descending order of intensity. Canonical sEV marker proteins are highlighted in each plot. D Number of proteins identified in the two groups. Data are presented as mean ± SD. Statistical significance was assessed using the Student’s t -test (ns, not significant). E Total protein intensity in the two groups. Data are presented as median (bold horizontal line) with interquartile range (box). Statistical significance was assessed using the Mann–Whitney U -test (***, P < 0.001). F t-SNE plot showing clustering of proteomic profiles from eight samples. G Scatter plot of 915 proteins identified in Ishikawa-sEVs, displaying protein intensities in I-Ishikawa-sEVs against P-Ishikawa-sEVs. DEPs are colour-coded: red indicates upregulation and blue indicates downregulation in I-Ishikawa-sEVs compared with P-Ishikawa-sEVs. The labelled proteins are commonly differentially expressed in both UF-sEVs and Ishikawa-sEVs during the implantation phase. H Dot plots showing the expression levels of galectin-3 in each sample. Data are presented as mean ± SD. Statistical significance was assessed using the Mann–Whitney U -test (*, P < 0.05; **, P < 0.01)). Abbreviations: FBS, Foetal bovine serum; E 2 , β-estradiol; P 4 , Progesterone; <t>cAMP,</t> <t>8-bromoadenosine</t> <t>3′,5′-cyclic</t> monophosphate; P-Ishikawa-sEVs, Proliferative-phase Ishikawa-sEVs; I-Ishikawa-sEVs, Implantation-phase Ishikawa-sEVs; DEP, Differentially expressed protein
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Proteomic analysis of Ishikawa-sEVs. A Timeline of the experimental protocol for hormonal treatment of Ishikawa cells. B Concentration of sEVs (µg/mL; total protein amount of sEVs per volume of conditioned culture medium) in the two groups. Data are presented as mean ± SD. Statistical significance was assessed using the Student’s t -test (ns, not significant). C Dot plots showing the expression levels of proteins identified in Ishikawa-sEVs (915 proteins) ranked in descending order of intensity. Canonical sEV marker proteins are highlighted in each plot. D Number of proteins identified in the two groups. Data are presented as mean ± SD. Statistical significance was assessed using the Student’s t -test (ns, not significant). E Total protein intensity in the two groups. Data are presented as median (bold horizontal line) with interquartile range (box). Statistical significance was assessed using the Mann–Whitney U -test (***, P < 0.001). F t-SNE plot showing clustering of proteomic profiles from eight samples. G Scatter plot of 915 proteins identified in Ishikawa-sEVs, displaying protein intensities in I-Ishikawa-sEVs against P-Ishikawa-sEVs. DEPs are colour-coded: red indicates upregulation and blue indicates downregulation in I-Ishikawa-sEVs compared with P-Ishikawa-sEVs. The labelled proteins are commonly differentially expressed in both UF-sEVs and Ishikawa-sEVs during the implantation phase. H Dot plots showing the expression levels of galectin-3 in each sample. Data are presented as mean ± SD. Statistical significance was assessed using the Mann–Whitney U -test (*, P < 0.05; **, P < 0.01)). Abbreviations: FBS, Foetal bovine serum; E 2 , β-estradiol; P 4 , Progesterone; <t>cAMP,</t> <t>8-bromoadenosine</t> <t>3′,5′-cyclic</t> monophosphate; P-Ishikawa-sEVs, Proliferative-phase Ishikawa-sEVs; I-Ishikawa-sEVs, Implantation-phase Ishikawa-sEVs; DEP, Differentially expressed protein
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Proteomic analysis of Ishikawa-sEVs. A Timeline of the experimental protocol for hormonal treatment of Ishikawa cells. B Concentration of sEVs (µg/mL; total protein amount of sEVs per volume of conditioned culture medium) in the two groups. Data are presented as mean ± SD. Statistical significance was assessed using the Student’s t -test (ns, not significant). C Dot plots showing the expression levels of proteins identified in Ishikawa-sEVs (915 proteins) ranked in descending order of intensity. Canonical sEV marker proteins are highlighted in each plot. D Number of proteins identified in the two groups. Data are presented as mean ± SD. Statistical significance was assessed using the Student’s t -test (ns, not significant). E Total protein intensity in the two groups. Data are presented as median (bold horizontal line) with interquartile range (box). Statistical significance was assessed using the Mann–Whitney U -test (***, P < 0.001). F t-SNE plot showing clustering of proteomic profiles from eight samples. G Scatter plot of 915 proteins identified in Ishikawa-sEVs, displaying protein intensities in I-Ishikawa-sEVs against P-Ishikawa-sEVs. DEPs are colour-coded: red indicates upregulation and blue indicates downregulation in I-Ishikawa-sEVs compared with P-Ishikawa-sEVs. The labelled proteins are commonly differentially expressed in both UF-sEVs and Ishikawa-sEVs during the implantation phase. H Dot plots showing the expression levels of galectin-3 in each sample. Data are presented as mean ± SD. Statistical significance was assessed using the Mann–Whitney U -test (*, P < 0.05; **, P < 0.01)). Abbreviations: FBS, Foetal bovine serum; E 2 , β-estradiol; P 4 , Progesterone; <t>cAMP,</t> <t>8-bromoadenosine</t> <t>3′,5′-cyclic</t> monophosphate; P-Ishikawa-sEVs, Proliferative-phase Ishikawa-sEVs; I-Ishikawa-sEVs, Implantation-phase Ishikawa-sEVs; DEP, Differentially expressed protein
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Proteomic analysis of Ishikawa-sEVs. A Timeline of the experimental protocol for hormonal treatment of Ishikawa cells. B Concentration of sEVs (µg/mL; total protein amount of sEVs per volume of conditioned culture medium) in the two groups. Data are presented as mean ± SD. Statistical significance was assessed using the Student’s t -test (ns, not significant). C Dot plots showing the expression levels of proteins identified in Ishikawa-sEVs (915 proteins) ranked in descending order of intensity. Canonical sEV marker proteins are highlighted in each plot. D Number of proteins identified in the two groups. Data are presented as mean ± SD. Statistical significance was assessed using the Student’s t -test (ns, not significant). E Total protein intensity in the two groups. Data are presented as median (bold horizontal line) with interquartile range (box). Statistical significance was assessed using the Mann–Whitney U -test (***, P < 0.001). F t-SNE plot showing clustering of proteomic profiles from eight samples. G Scatter plot of 915 proteins identified in Ishikawa-sEVs, displaying protein intensities in I-Ishikawa-sEVs against P-Ishikawa-sEVs. DEPs are colour-coded: red indicates upregulation and blue indicates downregulation in I-Ishikawa-sEVs compared with P-Ishikawa-sEVs. The labelled proteins are commonly differentially expressed in both UF-sEVs and Ishikawa-sEVs during the implantation phase. H Dot plots showing the expression levels of galectin-3 in each sample. Data are presented as mean ± SD. Statistical significance was assessed using the Mann–Whitney U -test (*, P < 0.05; **, P < 0.01)). Abbreviations: FBS, Foetal bovine serum; E 2 , β-estradiol; P 4 , Progesterone; cAMP, 8-bromoadenosine 3′,5′-cyclic monophosphate; P-Ishikawa-sEVs, Proliferative-phase Ishikawa-sEVs; I-Ishikawa-sEVs, Implantation-phase Ishikawa-sEVs; DEP, Differentially expressed protein

Journal: Cell Communication and Signaling : CCS

Article Title: Galectin-3 in endometrial small extracellular vesicles promotes cytotrophoblast cell fusion during the implantation phase

doi: 10.1186/s12964-025-02586-1

Figure Lengend Snippet: Proteomic analysis of Ishikawa-sEVs. A Timeline of the experimental protocol for hormonal treatment of Ishikawa cells. B Concentration of sEVs (µg/mL; total protein amount of sEVs per volume of conditioned culture medium) in the two groups. Data are presented as mean ± SD. Statistical significance was assessed using the Student’s t -test (ns, not significant). C Dot plots showing the expression levels of proteins identified in Ishikawa-sEVs (915 proteins) ranked in descending order of intensity. Canonical sEV marker proteins are highlighted in each plot. D Number of proteins identified in the two groups. Data are presented as mean ± SD. Statistical significance was assessed using the Student’s t -test (ns, not significant). E Total protein intensity in the two groups. Data are presented as median (bold horizontal line) with interquartile range (box). Statistical significance was assessed using the Mann–Whitney U -test (***, P < 0.001). F t-SNE plot showing clustering of proteomic profiles from eight samples. G Scatter plot of 915 proteins identified in Ishikawa-sEVs, displaying protein intensities in I-Ishikawa-sEVs against P-Ishikawa-sEVs. DEPs are colour-coded: red indicates upregulation and blue indicates downregulation in I-Ishikawa-sEVs compared with P-Ishikawa-sEVs. The labelled proteins are commonly differentially expressed in both UF-sEVs and Ishikawa-sEVs during the implantation phase. H Dot plots showing the expression levels of galectin-3 in each sample. Data are presented as mean ± SD. Statistical significance was assessed using the Mann–Whitney U -test (*, P < 0.05; **, P < 0.01)). Abbreviations: FBS, Foetal bovine serum; E 2 , β-estradiol; P 4 , Progesterone; cAMP, 8-bromoadenosine 3′,5′-cyclic monophosphate; P-Ishikawa-sEVs, Proliferative-phase Ishikawa-sEVs; I-Ishikawa-sEVs, Implantation-phase Ishikawa-sEVs; DEP, Differentially expressed protein

Article Snippet: Subsequently, the cells were divided into two groups: a proliferative-phase Ishikawa cell group (P-Ishikawa), cultured in basal medium with 10 –8 M E 2 , and an implantation-phase Ishikawa cell group (I-Ishikawa), cultured in basal medium with 10 –8 M E 2 , 10 –7 M progesterone (P 4 ; #P8783-1G, Sigma-Aldrich), 2.5 × 10 –4 M 8-bromoadenosine 3′,5′-cyclic monophosphate (cAMP; #05450-02, Nacalai Tesque), and 10 –5 M XAV939 (#247–00951, FUJIFILM Wako, Osaka, Japan).

Techniques: Concentration Assay, Expressing, Marker, MANN-WHITNEY