Journal: Journal of Translational Medicine

Article Title: Single-cell transcriptomic landscape of the mid-secretory eutopic endometrium reveals receptivity defects in adenomyosis

doi: 10.1186/s12967-026-07866-z

Figure Lengend Snippet: Characterization of eutopic endometrial epithelial cells in adenomyosis (AM). ( A ) UMAP plot showing four epithelial cell subclusters (Epi1–Epi4) ( B ) Heatmap of the top marker genes for each epithelial subcluster. ( C–E ) Gene Ontology (GO) enrichment analyses of top marker genes in Epi1 ( C ), Epi2 ( D ), and Epi3 ( E ). ( F ) Box plots showing the relative proportions of AM and control cells within each epithelial subcluster. ( G ) LGR5 and SOX9 mRNA expressions in mid-secretory eutopic endometrium from AM and control patients examined by qPCR analysis ( n = 12 per group). ( H , I ) Immunofluorescence staining for the expression and localization of LGR5 and SOX9. Scale bars, 50 μm; n = 12 per group. ( J ) Immunohistochemical staining for the expression of proliferation marker Ki-67. Scale bars, 50 μm; n = 12 per group. ( K ) Cell proliferation following plasmid LGR5 and SOX9 (p(LGR5 + SOX9)) overexpression in human endometrial epithelial cells (HEECs) isolated from mid-secretory endometrial tissues of controls ( n = 3) measured by CCK-8 assay. ( L ) Proliferation of HEECs transfected with p(LGR5 + SOX9) determined via EdU assay. Actively proliferating cells are visualized by green fluorescence, with all nuclei counterstained in blue. Scale bar = 100 μm; n = 3. ( M ) RT-qPCR quantification of proliferation markers ( MKI67 , PCNA ) and differentiation markers ( LIF , HOXA10 ) levels after LGR5 and SOX9 overexpression in control HEECs ( n = 3). ( N ) Representative immunofluorescence images for Ki-67 (red) and corresponding quantification of positive cells in HEECs. Scale bar = 100 μm; n = 3. ( O ) Western blotting of LIF and PCNA protein levels in HEECs transfected with p(LGR5 + SOX9). Data are presented as the mean ± SD, * P < 0.05, ** P < 0.01

Article Snippet: Control, LGR5, SOX9, and DIO2 overexpression plasmids (GeneCopoeia, China) were introduced using X-tremeGENE 9 (Roche, USA).

Techniques: Marker, Control, Immunofluorescence, Staining, Expressing, Immunohistochemical staining, Plasmid Preparation, Over Expression, Isolation, CCK-8 Assay, Transfection, EdU Assay, Fluorescence, Quantitative RT-PCR, Western Blot

Journal: Cell Transplantation

Article Title: Collagen-hyaluronic acid hydrogel with embedded chondrocytes as a platform for modeling early stages of endochondral ossification in vitro

doi: 10.1177/09636897251409464

Figure Lengend Snippet: Relative expression of chondrogenic genes during osteogenic differentiation: (a) SRY-Box Transcription Factor 9 ( SOX9 ) and (b) aggrecan ( ACAN ) at days 4, 7, 14, 21, and 28 in the studied hydrogels Col I, Col I/II-HA, Col I/II-HA+Cho, and Col I/II-HA+Cho_only. Mean of 2 −ΔΔCt + SD (n = 3). Expression compared to day 0 of the corresponding hydrogel, set as 1 (not shown on the graph). Two-way ANOVA with Tukey’s multiple comparisons test ( P ≤ 0.05). Comparisons for each time interval separately; significance: $ in comparison with Col I/II-HA, # in comparison with Col I/II-HA+Cho, and * in comparison with Col I/II-HA+Cho_only.

Article Snippet: For osteogenic cell differentiation, the following genes were examined: Col I ( COL1A1 gene, Hs00164004_m1), an early differentiation marker; alkaline phosphatase ( ALPL gene, Hs01029144_m1), an intermediate marker; osteocalcin ( BGLAP gene, Hs00165814_m1) and osteopontin ( SPP1 gene, Hs00959010_m1), late differentiation markers.

Techniques: Expressing, Comparison

Journal: Bioactive Materials

Article Title: Bioinspired scaffold recapitulating chondrogenic ontogeny and microenvironment for functional cartilage regeneration

doi: 10.1016/j.bioactmat.2025.11.041

Figure Lengend Snippet: Biological functional analysis of tri-layer scaffold systems. A) Representative images of migrated cells stained with crystal violet at 48 h. B) Number of migrating cells in control and CE-SKP. C) Glycosaminoglycan concentrations in control and CE-SKP at 7 and 14 days. D) qPCR analysis for COL2, SOX9, and ACAN. E) Western blotting of COL2 and SOX9. F) Alizarin red staining of control and CPH. G) ALP activity of control and CPH. H) qPCR analysis for COL1, COL10, OCN, and RUNX2. I) Western blotting of COL10, COL1, and RUNX2. J) Representative images of migrated cells stained with crystal violet at 48 h. K) Number of migrating cells in control and P2G3. L) Scanning electron microscope micrographs of P2G3. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

Article Snippet: After blocking the membranes for an hour at room temperature using fast blocking solution, the membranes were incubated with primary antibodies specific for SOX9 (1:2000, 67439-1-Ig, Proteintech), COL2 (1:200, NBP1-91056, Novus Biologicals), COL10 (1:500, 26984-1-AP, Proteintech), COL1 (1:2000, 67288-1-Ig, Proteintech), RUNX2 (1:200, 20700-1-AP, Proteintech), FGF2 (1:500, 11234-1-AP, Proteintech), FGF18 (1:500, 60341-1-Ig, Proteintech) and GAPDH (1:50000, 60004-1-Ig, Proteintech) for an entire night at 4 °C.

Techniques: Functional Assay, Staining, Control, Western Blot, Activity Assay, Microscopy

Journal: Journal of Translational Medicine

Article Title: Single-cell transcriptomic landscape of the mid-secretory eutopic endometrium reveals receptivity defects in adenomyosis

doi: 10.1186/s12967-026-07866-z

Figure Lengend Snippet: Identification, functional characterization, and trajectory analysis of stromal cell subclusters in adenomyosis (AM). ( A ) UMAP visualization of stromal subclusters (Str1–Str4). ( B ) Dot plots depicting the average expression of established markers indicated ecotype. ( C ) Pseudotime trajectory showing the progression of Str1, Str2, Str3 and Str4. ( D ) Gene Ontology (GO) terms significantly enriched across gene clusters with distinct pseudo-temporal patterns. ( E ) The distribution of DIO2 , PGR , and WNT5A in stromal cell subclusters by UMAP. ( F ) Violin plot of inflammatory scores across stromal subclusters. ( G ) Box plots of the relative proportions of AM and control cells in each stromal subcluster. ( H ) Separate pseudotime trajectories of stromal subclusters in control and AM groups

Article Snippet: Control, LGR5, SOX9, and DIO2 overexpression plasmids (GeneCopoeia, China) were introduced using X-tremeGENE 9 (Roche, USA).

Techniques: Functional Assay, Expressing, Control

Journal: Journal of Translational Medicine

Article Title: Single-cell transcriptomic landscape of the mid-secretory eutopic endometrium reveals receptivity defects in adenomyosis

doi: 10.1186/s12967-026-07866-z

Figure Lengend Snippet: DIO2 ⁺ Str4 stromal cells exhibit SASP-like activity and drive decidualization. ( A , B ) Representative immunohistochemistry (IHC) images of DIO2, CXCL12, IGFBP3, and MMP14 in mid-secretory eutopic endometrial tissues from controls and adenomyosis (AM) patients ( n = 12). Scale bars, 50 μm. ( C ) ELISA quantification of decidualization markers IGFBP1 and PRL in control and AM tissues ( n = 12). ( D ) Senescence-associated β-galactosidase (SA-β-gal) staining in control HESCs undergoing in vitro decidualization. ( E ) RT-qPCR analysis of SASP-related cytokines CXCL14, TIMP3, and IL15 in decidualized control HESCs ( n = 3). ( F ) ELISA measurement of secreted SASP cytokines in culture supernatant ( n = 3). ( G – K ) Effects of DIO2 knockdown on β-gal activity ( G ), SASP cytokine mRNA expression ( H ), and secreted protein levels ( I – K ) in control HESCs ( n = 3). ( L , M ) F-actin filaments staining in control HESCs following DIO2 knockdown during in vitro decidualization, visualized with Alexa Fluor 555–conjugated phalloidin. Scale bar = 50 μm, n = 3. Data are presented as the mean ± SD, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Control, LGR5, SOX9, and DIO2 overexpression plasmids (GeneCopoeia, China) were introduced using X-tremeGENE 9 (Roche, USA).

Techniques: Activity Assay, Immunohistochemistry, Enzyme-linked Immunosorbent Assay, Control, Staining, In Vitro, Quantitative RT-PCR, Knockdown, Expressing