Journal: bioRxiv

Article Title: Impact of vitamin D deficiency on defective endometrial decidualization and the repressive role of vitamin D receptor (VDR) in the epigenomic network

doi: 10.1101/2025.11.14.688535

Figure Lengend Snippet: (A) VDR mRNA expression levels in T-HESCs treated with EPC compared to untreated controls. (B) Western blot analysis of VDR protein levels in T-HESCs over 72 hr of EPC treatment, with kidney lysates from two individual female C57BL6/J mice as a positive control. (C) Validation of siVDR knockdown efficiency by qRT-PCR. (D) PRL and (E) IGFBP1 mRNA expression in siNT- and siVDR-transfected cells during EPC treatment. (F) VDR overexpression in lentiviral transduced T-HESCs was confirmed by qRT-PCR, and pLenti-GFP vector transduced T-HESCs were used as a control. (G) PRL and (H) IGFBP1 mRNA expression in VDR-overexpressing cells during EPC treatment. Data are presented as mean ± SEM. Statistical significance: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: Taqman probes for human genes were used as Hs99999901_s1, Hs00236877_m1, Hs00168730_m1, and Hs00172113_m1 for 18 s, PRL , IGFBP1 , and VDR , respectively.

Techniques: Expressing, Western Blot, Positive Control, Biomarker Discovery, Knockdown, Quantitative RT-PCR, Transfection, Over Expression, Plasmid Preparation, Control

Journal: Advanced Science

Article Title: Deciphering the Role and Mechanism of Decidual Monocyte‐Derived Macrophage Infiltration in Obstetric Antiphospholipid Syndrome at Single‐Cell Resolution

doi: 10.1002/advs.202503480

Figure Lengend Snippet: Abnormal infiltration of monocyte‐derived CCR2 + macrophages in OAPS decidua and their pro‐inflammatory roles. A) Bar plot showing the proportions of MDMs in decidual immune cells from OAPS patients and HCs in scRNA‐seq data. B) FCM analysis of the proportion of CCR2 + macrophages in the decidua from OAPS patients (n = 11) compared with HCs (n = 18). C) Representative images for CCR2 + and CCR2 − macrophages from the decidua of OAPS patients and HCs under the transmission electron microscope. Scale bar: 2 µm. D) Bubble plot showing the functional gene expressions in different types of decidual myeloid cells based on scRNA‐seq data. E) Bar plot showing the GO terms of marker genes in decidual MDMs. F) Bar plot showing the KEGG terms of marker genes in decidual MDMs. G) GSEA indicates the term of inflammatory response is significantly enriched in decidual CCR2 + macrophages. H) Bar plot showing the relative expression levels of genes related to inflammatory factors in decidual CCR2 + and CCR2 − macrophages, as detected using RT‐qPCR (n = 3). I) Representative immunoblots and semi‐quantified results of Arginase 1, NF‐κB P65, and STAT3 expressions in decidual CCR2 + and CCR2 − macrophages from OAPS patients (n = 3). J) Representative immunofluorescence images for CD14 (green), CCR2 (red), IGFBP1 (croci), HLA‐G (cyan), and DAPI (blue) co‐staining in decidua from HCs. Scale bar: 1000 µm. K) Representative immunofluorescence images for CD14 (green), CCR2 (red), IGFBP1 (croci), HLA‐G (cyan), and DAPI (blue) co‐staining in decidua from OAPS patients. Scale bar: 2000 µm. L) Line chart showing the effects of medium supernatants from CCR2 + and CCR2 − macrophages on HTR‐8/SVneo proliferation, as measured by CCK‐8 (n = 3). M) Bar plot showing relative expression levels of CASP3 , MMP2 , and MMP9 genes in HTR‐8/SVneo treated with supernatants from CCR2 + and CCR2 − macrophages, as detected by RT‐qPCR (n = 3). Data in (A) and (B) are presented as mean ± SD and analyzed using Student's t ‐test. Data in (H) and (I) are presented as mean ± SD and analyzed using a paired t ‐test. Data in (L) and (M) are presented as mean ± SD and analyzed using one‐way ANOVA with Dunnett's multiple comparisons test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. OAPS, obstetric antiphospholipid syndrome; HCs, healthy controls; MDM, monocyte‐derived macrophage; FCM, flow cytometry; scRNA‐seq, single‐cell RNA sequencing; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes; GSEA, Gene Set Enrichment Analysis; RT‐qPCR, real‐time quantitative polymerase chain reaction; SD, standard deviation; ANOVA, analysis of variance.

Article Snippet: CD14 Mouse Monoclonal antibody (Proteintech, China, #60253‐1‐Ig, 1:500), IGFBP1 Rabbit Polyclonal Antibody (Proteintech, China, #13981‐1‐AP, 1:2000), CCR2b‐specific Polyclonal antibody (Proteintech, China, #16154‐1‐AP, 1:800), HLA‐G Mouse Monoclonal antibody (Proteintech, China, #66447‐1‐Ig, 1:800), MCP‐1 Rabbit Polyclonal antibody (Proteintech, China, #26161‐1‐AP, 1:500), vWF Rabbit Polyclonal Antibody (Proteintech, China, #27186‐1‐AP, 1:300), DARC Polyclonal antibody (Proteintech, China, #55185‐1‐AP, 1:200), F4/80 Polyclonal antibody (CST, USA, #70 076, 1:600) and CCR2a‐specific Polyclonal antibody (Proteintech, China, #16153‐1‐AP, 1:800) were used as primary antibodies.

Techniques: Derivative Assay, Transmission Assay, Microscopy, Functional Assay, Marker, Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Staining, CCK-8 Assay, Flow Cytometry, RNA Sequencing, Real-time Polymerase Chain Reaction, Standard Deviation

Journal: Journal of Pharmaceutical Analysis

Article Title: Prioritization of potential drug targets for diabetic kidney disease using integrative omics data mining and causal inference

doi: 10.1016/j.jpha.2025.101265

Figure Lengend Snippet: Differential expression of prioritized targets in tissue immunostaining. (A) Immunohistochemistry staining validating increased expression of insulin-like growth factor binding protein 4 (IGFBP4), family with sequence similarity 3 member C (FAM3C), prostaglandin D2 synthase (PTGDS), and insulin-like growth factor binding protein 1 (IGFBP1) in renal biopsy specimens from diabetic kidney disease (DKD) and healthy volunteers. (B) Periodic acid-Schiff (PAS), periodic-acid silver methenamine (PASM) staining, blood glucose and proteinuria confirming successful modeling of diabetic (NDKD) and DKD mice. (C) Immunohistochemistry identifying boosted expression of IGFBP1, FAM3C, and PTGDS in DKD mice. (D) Representative immunofluorescence indicating elevated expression of FAM3C (red), IGFBP4 (green), and PTGDS (yellow) in the renal tissues of DKD mice across different groups. Nuclei were counterstained with 4,6-diamidino-2-phenylindole (DAPI) (blue). ∗∗∗ P < 0.001, ∗∗ P < 0.01, ∗ P < 0.05. IRS: immunoreactivity score.

Article Snippet: The specific primary antibodies included: family with sequence similarity 3 member C (FAM3C) antibody (HUABIO, Hangzhou, China, ER1908-63, 1:200), insulin-like growth factor binding protein 1 (IGFBP1) antibody (Proteintech, Wuhan, China, 13981-1-AP, 1:2000), prostaglandin D2 synthase (PTGDS) antibody (Proteintech, Wuhan, China, 10754-2-AP, 1:1000) and neuroblastoma suppressor of tumorigenicity 1 (NBL1) antibody (Affinity Biosciences, Nanjing, China, DF3177, 1:200).

Techniques: Quantitative Proteomics, Immunostaining, Immunohistochemistry, Staining, Expressing, Binding Assay, Sequencing, Immunofluorescence

Journal: Biology of Reproduction

Article Title: The roles of AKT isoforms in decidualization and embryo implantation using a Progesterone Receptor-Cre mouse model ^†

doi: 10.1093/biolre/ioaf062

Figure Lengend Snippet: Induction of artificial decidualization in the PGR-Cre mouse model. (A) Schematic representation of the artificial decidualization protocol used in this study. (B) A representative uterus obtained from artificial decidualization induction. Female mice were mated with a vasectomized WT male. The uterus was harvested on day 8 of pseudopregnancy following sesame oil injection in the right uterine horn on day 4. The contralateral horn served as a control. Scale bar is 5 mm. (C) Validation of successful decidualization by physiology (H&E) and expression of a specific marker (IGFBP1). Paraffin-embedded control and oil-injected uteri from a WT mouse were sectioned transversally at a thickness of 6 μm. H&E staining was performed, and an IGFBP1 antibody was used to detect its expression by immunohistochemistry. Images were taken using a 40× microscope objective and a 5.1-MP camera. Scale bar represents 100 μm. GE, glandular epithelium; LE, luminal epithelium; S, stroma. (D) Representative decidualized uteri from each genotype. The percentage at the top of the image represents the success rate of artificial decidualization induction. Images were taken using a 32-MP camera. Scale bar is 5 mm. (E) The success rate of artificial decidualization induction was calculated for each genotype. Data were analyzed using a binomial test on GraphPad Prism 8 to compare each group to the expected rate from WT. A minimum of 18 independent experiments were performed ( n = 18 mice/genotype). **** p < 0.0001. GE, glandular epithelium; LE, luminal epithelium; S, stroma.

Article Snippet: The primary antibodies used from Cell Signaling Technology: IGFBP1 (#31025, 1:200), Progesterone Receptor A/B (#8757, 1:150), PTGS2 (COX2) (#12282, 1:300), AKT (#4691, 1:150), pAKT substrate (#9614, 1:500), FOXO1 (#2880, 1:100), NF-kappaB p65 (#8242, 1:200), and FOXA2 (#8186, 1:400).

Techniques: Injection, Control, Biomarker Discovery, Expressing, Marker, Staining, Immunohistochemistry, Microscopy