primary antibodies  (R&D Systems)

Journal: Reproduction & Fertility

Article Title: Aspirin enhances endometrial decidualization markers in vitro among women with and without endometriosis

doi: 10.1530/RAF-25-0034

Figure Lengend Snippet: (A, B, C, D) Aspirin (ASA) enhances decidualization markers of endometrial stromal cells (ESCs). Treatment of ESCs with ASA (1–2.5 mM) prior to stimulation with either (A) cAMP alone or (B) cAMP + MPA enhances decidualization markers, as determined by IGFBP1 protein levels by ELISA when compared to vehicle (Veh)-treated ESCs. Treatment of ESCs with ASA (1–2.5 mM) prior to stimulation with either (C) cAMP alone or (D) cAMP + MPA enhances decidualization markers, as determined by PRL protein levels by ELISA compared to vehicle (Veh)-treated ESCs. Each dot represents data from ESCs isolated from one participant. Significance was determined by the Kruskal–Wallis test with post hoc Dunn’s multiple comparison test; P -values are shown. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: After 48 h, culture supernatants were collected after brief centrifugation and cell-free supernatants were analyzed for decidualization markers, IGFBP1 or PRL, using Human IGFBP1 and PRL DuoSet ELISA Kits (R&D Systems®, USA), respectively, according to the manufacturer’s directions and as previously reported ( Warren et al. 2018 , Nayyar et al. 2020 , Delenko et al. 2024 ).

Techniques: Enzyme-linked Immunosorbent Assay, Isolation, Comparison

Journal: Advanced Science

Article Title: IGFBP5 Restores Endometrial Receptivity and Rescues Implantation Failure in Polycystic Ovary Syndrome

doi: 10.1002/advs.202520455

Figure Lengend Snippet: IL‐22 promotes IGFBP5 expression by activating the STAT3 signaling pathway. (A) Volcano plots showing DEGs in PCOS and PCOS+IL‐22 organoids (|log2FC| > 1, adj. P < 0.05), red dot represent for upregulated DEGs, blue dot represent for down‐regulated DEGs, n = 3 per group; (B) Immunohistochemistry staining of IGFBP5 in CON, DHEA and DHEA + IL‐22 mice uterus on day 4 of pregnancy; le, luminal epithelium; ge, glandular epithelium; s, stroma, scales: 50 µm; (C) representative immunoblots of CON ( n = 7), DHEA ( n = 3) and DHEA + IL‐22 ( n = 3) mice uterus on day 4 of pregnancy; (D) Spearman correlation analysis between relative IGFBP5 protein levels and pSTAT3/STAT3 levels in mice uterus on day 4 of pregnancy ( n = 13 in total). (E) qPCR results of IGFBP5 expression in STAT3‐OE plasmids‐transfected Ishikawa cells, n = 3 per group; (F) Representative immunofluorescence staining pictures of IGFBP5 in Ishikawa cells transfected with Vector or STAT3‐OE plasmids, scale bars: 50 µm; (G‐H) Binding motif (G) and predicted specific binding sites (H) of STAT3 and the IGFBP5 gene promoter; (I) Dual‐luciferase reporter assay of 293T cells, transfected with the indicated plasmids, n = 3 per group. For E and I, data are presented as mean ± SEM, the P value was determined by a two‐tailed Student's t ‐test. For B and F, the experiments were performed in three replicates.

Article Snippet: To clarify the amelioration of IGFBP5 on impaired endometrial receptivity in PCOS, recombinant human IGFBP5 (2 mg/kg, HY‐ P72378 , Med Chem Express) was intraperitoneally injected to DHEA‐treated mice daily from D1 to day 4 of pregnancy (D4), with PBS injected as a control in the CON and DHEA group [ , ].

Techniques: Expressing, Immunohistochemistry, Staining, Western Blot, Transfection, Immunofluorescence, Plasmid Preparation, Binding Assay, Luciferase, Reporter Assay, Two Tailed Test

Journal: Advanced Science

Article Title: IGFBP5 Restores Endometrial Receptivity and Rescues Implantation Failure in Polycystic Ovary Syndrome

doi: 10.1002/advs.202520455

Figure Lengend Snippet: IL‐22 acts through IGFBP5 to alleviate implantation disorders in PCOS. (A) Experiment design for mice treatment and intrauterine injection of lentivirus; (B) Images presenting implantation sites (ISs) in three groups; (C) Number of ISs in three groups, n = 5 per group; (D) Percentage of mice with ISs in mice that had a vaginal plug; (E) Representative immunofluorescence staining of MUC1 and immunohistochemistry staining of Ki‐67 in mice uterus on day 4 of pregnancy, n = 5 per group, scale bars: 100 µm; (F) Relative gene expression of Esr1 and Pgr in mice uterus on day 4 of pregnancy, n = 5 per group; (G‐H) Representative immunohistochemistry staining images (G) and quantitative results (H) of ERα and PR protein in mice uterus on day 4 of pregnancy, n = 5 per group, scale bars: 100 µm; (I‐J) Relative gene expression of estrogen responsive genes Lif and Muc1 (I) and progesterone responsive genes Hand2 , Hoxa10 , Areg and Ihh (J) in mice uterus on day 4 of pregnancy, n = 5 per group. For C, data is presented as medians with interquartile ranges, and the P value was determined by Kruskal–Wallis test followed by Dunn's post hoc test. For F, H‐J, data are presented as mean ± SEM, the P value was determined by one‐way ANOVA with Tukey's multiple comparison post hoc test. For E and G, the experiments were performed in three replicates; le, luminal epithelium; ge, glandular epithelium; s, stroma.

Article Snippet: To clarify the amelioration of IGFBP5 on impaired endometrial receptivity in PCOS, recombinant human IGFBP5 (2 mg/kg, HY‐ P72378 , Med Chem Express) was intraperitoneally injected to DHEA‐treated mice daily from D1 to day 4 of pregnancy (D4), with PBS injected as a control in the CON and DHEA group [ , ].

Techniques: Injection, Immunofluorescence, Staining, Immunohistochemistry, Gene Expression, Comparison

Journal: Advanced Science

Article Title: IGFBP5 Restores Endometrial Receptivity and Rescues Implantation Failure in Polycystic Ovary Syndrome

doi: 10.1002/advs.202520455

Figure Lengend Snippet: IGFBP5 supplementation improves endometrial receptivity in PCOS‐derived endometrium. (A) Relative expression of IGFBP5 in proliferative and secretory endometrium of control women, n = 8 per group; (B) Representative immunohistochemistry staining of IGFBP5 in human endometrium of proliferative and secretory phase of control women, scale bars: 50 µm; (C) Quantitative analysis of IGFBP5 protein levels in epithelial and stromal cells of proliferative and secretory endometrium of control women, n = 3 per group; (D) Relative expression of IGFBP5 in secretory endometrium from the CON and PCOS groups, n = 8 per group; (E‐F) Representative immunohistochemistry staining (E) and its quantitative analysis (F) of IGFBP5 protein levels in the secretory endometrium from the CON and PCOS groups, n = 6 per group, scale bars: 50 µm; (G) IGFBP5 mRNA levels in DHT‐treated Ishikawa cells, n = 3 per group; (H) The percentage of JAr spheroids that adhere to Ishikawa cells treated with DHT (0.5 µM), DHT + IGFBP5 (100 ng/ml), DHT + IGF1 (50 ng/ml), DHT + IGFBP5 + IGF1 and DHT + IGFBP5 + NVP‐AEW541 (10 µM), n = 3 per group. For A, C, D, F, G, and H, data are presented as mean ± SEM. For A, C, D, and F, the P value was determined by a two‐tailed Student's t ‐test; for G and H, the P value was determined by one‐way ANOVA with Tukey's multiple comparison post hoc test. For B and E, the experiments were performed in three replicates; le, luminal epithelium; ge, glandular epithelium; s, stroma.

Article Snippet: To clarify the amelioration of IGFBP5 on impaired endometrial receptivity in PCOS, recombinant human IGFBP5 (2 mg/kg, HY‐ P72378 , Med Chem Express) was intraperitoneally injected to DHEA‐treated mice daily from D1 to day 4 of pregnancy (D4), with PBS injected as a control in the CON and DHEA group [ , ].

Techniques: Derivative Assay, Expressing, Control, Immunohistochemistry, Staining, Two Tailed Test, Comparison

Journal: Advanced Science

Article Title: IGFBP5 Restores Endometrial Receptivity and Rescues Implantation Failure in Polycystic Ovary Syndrome

doi: 10.1002/advs.202520455

Figure Lengend Snippet: IGFBP5 alleviates the implantation disorder of PCOS‐like mice. (A) Experiment design for mice treatment; (B) Images presenting ISs in CON, DHEA and DHEA +IGFBP5 mice; (C) Number of ISs in CON, DHEA and DHEA + IL‐22 mice, n = 5 per group; (D) Percentage of mice with ISs in mice that had a vaginal plug; (E‐F) Representative immunofluorescence staining of MUC1 (E) and immunohistochemistry staining of Ki‐67 (F) in mice uterus on day 4 of pregnancy, scale bars: 100 µm; (G) Relative gene expression of Esr1 and Pgr in mice uterus on day 4 of pregnancy, n = 5 per group; (H‐I) Representative immunohistochemistry staining (H) and quantitative results (I) of ERα and PR protein in mice uterus on day 4 of pregnancy, scale bars: 100 µm, n = 5 per group; (J‐K) Relative gene expression of estrogen responsive genes Lif and Muc1 (J) and progesterone responsive genes Hand2 , Hoxa10 , Areg and Ihh (K) in mice uterus on day 4 of pregnancy, n = 5 per group. For C, data is presented as medians with interquartile ranges, the P value was determined by Kruskal–Wallis test followed by Dunn's post hoc test. For G and I‐K, data are presented as mean ± SEM, the P value was determined by one‐way ANOVA with Tukey's multiple comparison post hoc test. For E, F, and H, the experiments were performed in three replicates; le, luminal epithelium; ge, glandular epithelium; s, stroma.

Article Snippet: To clarify the amelioration of IGFBP5 on impaired endometrial receptivity in PCOS, recombinant human IGFBP5 (2 mg/kg, HY‐ P72378 , Med Chem Express) was intraperitoneally injected to DHEA‐treated mice daily from D1 to day 4 of pregnancy (D4), with PBS injected as a control in the CON and DHEA group [ , ].

Techniques: Immunofluorescence, Staining, Immunohistochemistry, Gene Expression, Comparison