Journal: Experimental and Therapeutic Medicine

Article Title: MUC21 induces the viability and migration of glioblastoma via the STAT3/AKT pathway

doi: 10.3892/etm.2022.11260

Figure Lengend Snippet: Relationship between MUC21 expression and clinicopathological characteristics of patients with glioblastoma (n=47).

Article Snippet: Slides were then incubated with MUC21 antibody (1:100; cat. no. NBP3-06591; Novus Biologicals, LLC) for 2 h at room temperature.

Techniques: Expressing

Journal: Experimental and Therapeutic Medicine

Article Title: MUC21 induces the viability and migration of glioblastoma via the STAT3/AKT pathway

doi: 10.3892/etm.2022.11260

Figure Lengend Snippet: MUC21 is highly expressed in human GBM tissues. (A) A reverse transcription-quantitative PCR assay were conducted to determine the mRNA levels of MUC21 in GBM and corresponding adjacent non-cancerous tissues. A paired Student's t-test was used for the analysis of tumor and adjacent non-tumor samples. (B) Western blotting was performed to determine MUC21 expression in three GBM cell lines. (C) Immunohistochemistry was performed to evaluate the protein levels of MUC21 in GBM tumor and corresponding adjacent non-cancerous tissues. Magnification, x100 and x200 (scale bar, 5 mm). * P<0.05 as indicated. GBM glioblastoma; MUC21, mucin 21.

Article Snippet: Slides were then incubated with MUC21 antibody (1:100; cat. no. NBP3-06591; Novus Biologicals, LLC) for 2 h at room temperature.

Techniques: Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Immunohistochemistry

Journal: Experimental and Therapeutic Medicine

Article Title: MUC21 induces the viability and migration of glioblastoma via the STAT3/AKT pathway

doi: 10.3892/etm.2022.11260

Figure Lengend Snippet: MUC21 expression is downregulated in U251 and U87 cells after MUC21 shRNA transfection. (A) Reverse transcription-quantitative PCR assays were performed to measure MUC21 mRNA levels in U251 and U87 cells following control or MUC21 shRNA transfection. (B) Western blot analysis was performed to determine the protein expression of MUC21 in U251 and U87 cells following control or MUC21 shRNA transfection. * P<0.05 vs. shRNA. MUC21, mucin 21; NC, negative control; shRNA, short hairpin RNA.

Article Snippet: Slides were then incubated with MUC21 antibody (1:100; cat. no. NBP3-06591; Novus Biologicals, LLC) for 2 h at room temperature.

Techniques: Expressing, shRNA, Transfection, Reverse Transcription, Real-time Polymerase Chain Reaction, Control, Western Blot, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: MUC21 induces the viability and migration of glioblastoma via the STAT3/AKT pathway

doi: 10.3892/etm.2022.11260

Figure Lengend Snippet: MUC21 promotes GBM cell viability and migration in vitro . (A) Cell viability was assessed after MUC21 depletion in U87 and U251 through colony formation assays. (B) MTT assays were performed to detect the viability of cells transfected with the indicated shRNA. (C) Wound closure assay of GBM cells transfected with the indicated shRNA were performed for the quantification of migrated cells after 24 h. Magnification, x200. Western blotting assays were performed to analyze the expression of (D) Ki-67 and PCNA, and (E) MMP2 and MMP9 in GBM cells transfected with the indicated shRNA. Relative expression was analyzed. * P<0.05 and ** P<0.01 vs. the shNC group. GBM glioblastoma; MUC21, mucin 21; PCNA, proliferating cell nuclear antigen; shRNA, short hairpin RNA; NC, negative control.

Article Snippet: Slides were then incubated with MUC21 antibody (1:100; cat. no. NBP3-06591; Novus Biologicals, LLC) for 2 h at room temperature.

Techniques: Migration, In Vitro, Transfection, shRNA, Wound Closure Assay, Western Blot, Expressing, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: MUC21 induces the viability and migration of glioblastoma via the STAT3/AKT pathway

doi: 10.3892/etm.2022.11260

Figure Lengend Snippet: MUC21 contributes to GBM progression via the STAT3 and AKT signaling pathway. (A) The protein levels of p-STAT3, STAT3, p-AKT and AKT in shMUC21-transfected or control U251 cells were detected by immunoblot assays. (B) The protein levels of MUC21, p-STAT3, STAT3, p-AKT and AKT in MUC21-overexpressing or control U251 cells were detected by immunoblot assays. The relative expression was analyzed. * P<0.05 and ** P<0.01 vs. shNC or Con. Con, control; MUC21, mucin 21; NC, negative control; p-, phosphorylated; shRNA, short hairpin RNA.

Article Snippet: Slides were then incubated with MUC21 antibody (1:100; cat. no. NBP3-06591; Novus Biologicals, LLC) for 2 h at room temperature.

Techniques: Transfection, Control, Western Blot, Expressing, Negative Control, shRNA

Journal: Experimental and Therapeutic Medicine

Article Title: MUC21 induces the viability and migration of glioblastoma via the STAT3/AKT pathway

doi: 10.3892/etm.2022.11260

Figure Lengend Snippet: MUC21 stimulates glioblastoma progression through the STAT3 and AKT pathway in vivo . (A) U251 cells stably transfected with control or MUC21 shRNA vectors were subcutaneously implanted into nude mice. Tumor volume was monitored every week. Isolated tumors were displayed in (A). Tumor growth curves were compared between control and MUC21 depletion groups. Murine weight was also measured. (B) An immunohistochemistry assay was conducted to assess MUC21 protein levels in control or MUC21-depleted groups (scale bar, 200 µm). (C) Western blotting assays were performed to assess the expression levels of the indicated proteins in control or MUC21-depleted tumor tissues isolated from nude mice. * P<0.05 and ** P<0.01 vs. shNC. MUC21, mucin 21; NC, negative control; PCNA, proliferating cell nuclear antigen; p-, phosphorylated; shRNA, short hairpin RNA.

Article Snippet: Slides were then incubated with MUC21 antibody (1:100; cat. no. NBP3-06591; Novus Biologicals, LLC) for 2 h at room temperature.

Techniques: In Vivo, Stable Transfection, Transfection, Control, shRNA, Isolation, Immunohistochemistry, Western Blot, Expressing, Negative Control

Journal: Journal of Oral and Maxillofacial Surgery, Medicine, and Pathology

Article Title: Searching for new early detection markers of OED and oral SCC using oral liquid-based cytology

doi: 10.1016/j.ajoms.2023.11.007

Figure Lengend Snippet: Fig. 1. Papanicolaou staining in liquid-based cytology (LBC) samples and BRD4, c-MYC, TP53, and MUC21 immunohistochemical study in oral cytological samples: (A) NILM, (B) LSIL, (C) HSIL, and (D) SCC. (E-H) BRD4 immunocytochemical staining. Although BRD4 staining was generally negative in (E) NILM samples, (F) positive nuclear staining was observed in LSIL, (G) HSIL, and (H) SCC samples. (I-L) c-MYC immunocytochemical staining. Although c-MYC staining was generally negative in (I) NILM samples, positive nuclear staining was observed in (J) LSIL, (K) HSIL, and (L) SCC samples. (M-P) TP53 immunocytochemical staining. Although TP53 staining was generally negative in (M) NILM, (N) LSIL, and (O) HSIL samples, positive nuclear staining was observed in (P) SCC samples. (Q-T) MUC21 immunocytochemical staining. Although MUC21 staining was generally positive in (M) NILM and (N) LSIL, negative cytoplasmic staining was observed in (O) HSIL and (T) SCC samples. Original magnification, 400 x. Scale bars, 20 µm. NILM, negative for intraepithelial lesion or malignancy; LSIL, low‑grade squamous intra epithelial lesion; HSIL, high‑grade squamous intraepithelial lesion; SCC, squamous cell carcinoma.

Article Snippet: All slides were subjected to antigen retrieval using 10 mM Tris-HCl, 1 mM EDTA-2Na (pH8.0) in a microwave oven of 1000 W for 20 min, followed by incubation with a rabbit polyclonal anti-TP53 antibody (1:50 dilution; clone ab131442; Abcam, Cambridge, MA, USA), a rabbit monoclonal anti-cMYC antibody (1:100 dilution; clone ab32072; Abcam), a rabbit polyclonal anti-MUC21 antibody (1:200 dilution; clone NBP2–31023; Novus), or a rabbit monoclonal anti-BRD4 antibody (1:100 dilution; clone ab128874; Abcam).

Techniques: Staining, Immunohistochemical staining

Journal: Journal of Oral and Maxillofacial Surgery, Medicine, and Pathology

Article Title: Searching for new early detection markers of OED and oral SCC using oral liquid-based cytology

doi: 10.1016/j.ajoms.2023.11.007

Figure Lengend Snippet: Fig. 2. Histopathological and immunohistochemical expressions patterns of BRD4, c‑MYC, TP53, and MUC21 in normal epithelium (NOE), hyperplasia (HYP), oral epithelial dysplasia (OED), and squamous cell carcinoma (SCC) samples. (A, E, I, M, Q) NOE, (B, F, J, N, R) HYP, (C, G, K, O, S) OED and (D, H, L, P, T) SCC. (A-D) Hematoxylin and eosin (H&E), (E-H) BRD4, (I-L) c-MYC, (M-P) TP53, and (Q-T) MUC21. Original magnification, 100x. Scale bars, 100 µm.

Article Snippet: All slides were subjected to antigen retrieval using 10 mM Tris-HCl, 1 mM EDTA-2Na (pH8.0) in a microwave oven of 1000 W for 20 min, followed by incubation with a rabbit polyclonal anti-TP53 antibody (1:50 dilution; clone ab131442; Abcam, Cambridge, MA, USA), a rabbit monoclonal anti-cMYC antibody (1:100 dilution; clone ab32072; Abcam), a rabbit polyclonal anti-MUC21 antibody (1:200 dilution; clone NBP2–31023; Novus), or a rabbit monoclonal anti-BRD4 antibody (1:100 dilution; clone ab128874; Abcam).

Techniques: Immunohistochemical staining

Journal: Journal of Oral and Maxillofacial Surgery, Medicine, and Pathology

Article Title: Searching for new early detection markers of OED and oral SCC using oral liquid-based cytology

doi: 10.1016/j.ajoms.2023.11.007

Figure Lengend Snippet: Fig. 4. The correlation between the labeling index and the relative mRNA levels of each marker (BRD4, c-MYC, TP53, and MUC21) in the oral cytological specimens. These markers displayed significant positive correlations: (A) BRD4 (R= 0.781, p < 0.01), (B) c‑MYC (R=0.807, p < 0.01), (C) TP53 (R=0.606, p < 0.01), and (D) MUC21 (R=0.514, p < 0.05).

Article Snippet: All slides were subjected to antigen retrieval using 10 mM Tris-HCl, 1 mM EDTA-2Na (pH8.0) in a microwave oven of 1000 W for 20 min, followed by incubation with a rabbit polyclonal anti-TP53 antibody (1:50 dilution; clone ab131442; Abcam, Cambridge, MA, USA), a rabbit monoclonal anti-cMYC antibody (1:100 dilution; clone ab32072; Abcam), a rabbit polyclonal anti-MUC21 antibody (1:200 dilution; clone NBP2–31023; Novus), or a rabbit monoclonal anti-BRD4 antibody (1:100 dilution; clone ab128874; Abcam).

Techniques: Labeling, Marker

Journal: Journal of Oral and Maxillofacial Surgery, Medicine, and Pathology

Article Title: Searching for new early detection markers of OED and oral SCC using oral liquid-based cytology

doi: 10.1016/j.ajoms.2023.11.007

Figure Lengend Snippet: Fig. 5. Receiver operating characteristic analysis for LSIL or higher category specimens were screened using BRD4 (red line), c-MYC (green line), MUC21 (brown line), and TP53 (blue line) as candidate markers. The optimal cut-off values of each markers were calculated using ‘closest-topleft’ (10, 28).

Article Snippet: All slides were subjected to antigen retrieval using 10 mM Tris-HCl, 1 mM EDTA-2Na (pH8.0) in a microwave oven of 1000 W for 20 min, followed by incubation with a rabbit polyclonal anti-TP53 antibody (1:50 dilution; clone ab131442; Abcam, Cambridge, MA, USA), a rabbit monoclonal anti-cMYC antibody (1:100 dilution; clone ab32072; Abcam), a rabbit polyclonal anti-MUC21 antibody (1:200 dilution; clone NBP2–31023; Novus), or a rabbit monoclonal anti-BRD4 antibody (1:100 dilution; clone ab128874; Abcam).

Techniques:

Journal: Science Advances

Article Title: Single-cell atlas of cervical organoids uncovers epithelial immune heterogeneity and intercellular cross-talk during Chlamydia infection

doi: 10.1126/sciadv.ady1640

Figure Lengend Snippet: ( A ) Immunofluorescence images of ectocervical (top) and endocervical (bottom) organoids, uninfected (left) or infected (right) for 48 hours with Chlamydia, stained for KRT5 (green), major outer membrane protein (MOMP) (red), KRT8 (gray), and DAPI (blue). ( B ) UMAP projection of single cells from ecto- and endocervical organoids, colored by infection status: uninfected (UI), infected (Inf), and bystander (Bstd). ( C and D ) UMAP showing reclustered ectocervical squamous epithelial population from (B), colored by infection status (C) and subtype identity (D). ( E ) Proportion of UI, Bstd, and Inf cells in each ectocervical squamous subtype. ( F to G ) UMAP showing reclustered endocervical columnar epithelia from (B), colored by infection status (F) and subtype (G). ( H ) Proportion of UI, Bstd, and Inf cells in each endocervical columnar subtype. ( I ) Heatmap of differentially regulated TFs between ecto- and endocervix across infection conditions; color bar depicts the TF activity scores from high (deep pink) to low (blue). ( J ) Violin plot of gene set enrichment scores for the GO term defense response to bacterium across epithelial compartments and infection states; statistical significance assessed by Wilcoxon rank-sum test with Holm-adjusted P values ( ****P ≤ 0.0001). ( K ) The relative expression of IFN-related genes across ecto- and endocervical subclusters; dot size represents the % of cells expressing a particular gene, and the color bar indicates the intensity of scaled mean expression levels ranging from high (red) to low (blue). ( L ) Gene-weighted density UMAP projections showing expression of STAT1 , STAT2 , and IRF9 across epithelial cells in (B). ( M ) Violin plot showing ISG15 expression across ecto- and endocervical organoids in uninfected, bystander, and infected states. ( N ) IHC images showing CDH1 (green), ISG15 (red), MOMP (gray), and DAPI (blue) in ecto- and endocervical organoids, uninfected (left) or infected (right). Yellow arrows mark infected cells; arrowheads indicate ISG15 + bystander cells.

Article Snippet: The following primary antibodies were used for immunofluorescence: mouse anti–acetylated tubulin–Alexa Fluor 647 (1:300, Santa Cruz Biotechnology, sc-23950-AF647), mouse anti–E-cadherin–Alexa Fluor 488 (1:50, BD Biosciences, 560061), mouse anti–E-cadherin (1:50, BD Biosciences, 610181), rabbit anti–KRT5–Alexa Fluor 488 (1:300, Abcam, ab193894), mouse anti-MUC5B (1:200, Abcam, ab77995), rabbit anti-MUC21 (1:200, ProteinAtlas, HPA052028), rabbit anti-KRT8 (1:200, Abcam, ab59400), mouse-anti-KRT6 (1:50, Abcam, ab18586), recombinant rabbit anti-PAX8 (1:200, Abcam, ab239363), goat anti– C. trachomatis major outer membrane protein (1:500, Bio-Rad, 1990-0804), rabbit anti–HLA-DQA1 antibody (EPR7300) (1:200, Abcam, ab128959), rabbit anti-ISG15 polyclonal antibody (1:200, Proteintech,15981-1-AP), and for labeling the DNA, 4′,6-diamidino-2-phenylindole (DAPI, Roche, 10236276001) were used.

Techniques: Immunofluorescence, Infection, Staining, Membrane, Activity Assay, Expressing

Journal: bioRxiv

Article Title: Single-cell atlas of patient-derived cervical organoids uncovers epithelial immune heterogeneity and intercellular crosstalk during Chlamydia infection

doi: 10.1101/2025.04.13.648603

Figure Lengend Snippet: a) Schematic of the experimental workflow modeling Chlamydia infection in patient-derived ecto- and endocervical organoids. b) Uniform Manifold Approximation and Projection (UMAP) of ecto- and endocervical epithelial cell clusters. Each dot represents a single cell color-coded by tissue type. c-d) UMAP projections highlighting squamous (c) and columnar (d) epithelial subclusters, with cells coloured by cluster. e) URD differentiation tree of ectocervical squamous epithelial cells; each dot represents a single cell, colored by subcluster. Cells ordered based on pseudotime values starting from early (top) to late (bottom). f) Gene expression dynamics of selected squamous markers along the pseudotime; lines represent expression trend of a particular gene. g) Differentiation trajectory of endocervical columnar epithelial cells, colored by subcluster and ordered by pseudotime (early to late). h) Expression dynamics of key columnar epithelial marker genes along pseudotime. i) UMAP visualization of organoid and tissue-derived cell clusters after data integration; cells are color-coded based on their dataset of origin. j) UMAP shows six major cell populations across the integrated dataset. k) UMAP depicting nine epithelial subclusters identified post-integration, with cells color-coded by cluster. l) Bar plot depicting the epithelial cell proportions from different datasets across integrated subclusters. m) Heatmap showing gene set enrichment scores for gene ontology (GO) biological processes across integrated epithelial clusters; scale bar denotes the z-scored enrichment values ranging from high (deep pink) to low (blue).

Article Snippet: The following primary antibodies were used for immunofluorescence: mouse anti-acetylated tubulin-Alexa-647 (Santa Cruz Biotechnology, sc-23950-AF647), mouse anti-E-cadherin-Alexa-488 (BD Biosciences, 560061), mouse anti-E-cadherin (BD Biosciences, 610181), rabbit anti-KRT5-Alexa488 (Abcam, ab193894 mouse anti-MUC5B (Abcam, ab77995), rabbit anti-MUC21 (ProteinAtlas, HPA052028), rabbit anti-KRT8 (1:200, Abcam, abab59400), mouse-anti-KRT6 (Abcam, ab18586), recombinant rabbit anti-PAX8 (Abcam, ab239363), goat anti- Chlamydia trachomatis Major Outer Membrane Protein (MOMP) (1:500, BIO-RAD, 1990-0804) and for labeling the DNA, DAPI (Roche, 10236276001) was used.

Techniques: Infection, Derivative Assay, Gene Expression, Expressing, Marker

Journal: bioRxiv

Article Title: Single-cell atlas of patient-derived cervical organoids uncovers epithelial immune heterogeneity and intercellular crosstalk during Chlamydia infection

doi: 10.1101/2025.04.13.648603

Figure Lengend Snippet: a) IHC images of ectocervix (upper panel) and endocervix (lower panel) organoids, either uninfected (left) or infected (right) with Chlamydia for 48 hours, stained with KRT5 (green), MOMP (red), KRT8 (gray). Nuclei are stained with DAPI (blue). b) UMAP projection of uninfected (UI), infected (Inf), and bystander (Bstd) epithelial cells from ecto- and endocervical organoids; each dot represents a single cell, colored by infection status. c-d) UMAP visualization of the re-clustered ectocervical squamous epithelial population from (b), colored by infection status (c) and subtypes (d). e) Bar plot showing the proportion of UI, Inf, and Bstd cells across squamous epithelial subclusters of the ectocervix. f-g) UMAP depicting re-clustered endocervical columnar epithelia from (b), colored by infection status (f) and subtypes (g). h) Bar plot shows the proportion of UI, Inf, and Bstd cells across columnar epithelial subclusters of endocervix. i) Heatmap of differentially regulated TFs between ecto- and endocervix across infection conditions; color bar depicts the TF activity scores from high (deep pink) to low (blue). j) Violin plot showing the gene set enrichment scores for GO term corresponding to defense response to bacterium across epithelial compartments and infection states. k) Heatmap showing hallmark pathway enrichment scores across epithelial subtypes and infection conditions. Columns represent individual cells color-coded by tissue and infection status. l) Dot plot showing the relative expression of interferon-related genes across ecto- and endocervical subclusters; circle size represents the percentage of cells expressing a particular gene, the color bar indicates the intensity of scaled mean expression levels ranging from high (red) to low (blue). m) Heatmap of TFs with highly variable activities across ecto- and endocervical samples and infection states, with subcluster annotations. n) Gene-weighted density UMAP projections showing expression of STAT1 , STAT2 , and IRF9 across epithelial cells in (b).

Article Snippet: The following primary antibodies were used for immunofluorescence: mouse anti-acetylated tubulin-Alexa-647 (Santa Cruz Biotechnology, sc-23950-AF647), mouse anti-E-cadherin-Alexa-488 (BD Biosciences, 560061), mouse anti-E-cadherin (BD Biosciences, 610181), rabbit anti-KRT5-Alexa488 (Abcam, ab193894 mouse anti-MUC5B (Abcam, ab77995), rabbit anti-MUC21 (ProteinAtlas, HPA052028), rabbit anti-KRT8 (1:200, Abcam, abab59400), mouse-anti-KRT6 (Abcam, ab18586), recombinant rabbit anti-PAX8 (Abcam, ab239363), goat anti- Chlamydia trachomatis Major Outer Membrane Protein (MOMP) (1:500, BIO-RAD, 1990-0804) and for labeling the DNA, DAPI (Roche, 10236276001) was used.

Techniques: Infection, Staining, Activity Assay, Expressing

Journal: bioRxiv

Article Title: Single-cell atlas of patient-derived cervical organoids uncovers epithelial immune heterogeneity and intercellular crosstalk during Chlamydia infection

doi: 10.1101/2025.04.13.648603

Figure Lengend Snippet: a) Dot plot showing mRNA profiles of PRRs, MHC genes, AMPs and cytokines across ecto- and endocervical epithelial subtypes and UI/Bstd/Inf conditions. The size of each dot reflects the percentage of cells expressing a specific gene, and the color bar signifies the intensity of scaled mean expression levels, ranging from high (red) to low (blue). b-c) qRT-PCR analysis of MHC genes ( HLA-B, HLA-C, HLA-DMB, HLA-DRA ) (b); PRRs ( TLR1, TLR2, TLR5 ) (c) expression in uninfected and infected ecto- and endocervical organoids at 48hpi. d) qRT-PCR analysis of DEFB1 expression in FACS sorted uninfected, infected, and bystander epithelial cells after 36hpi. Data represent mean ± s.d. from three technical replicates normalized to ectocervix control, and p-values were calculated using Student’s T-test. e) Infectivity assay of Chlamydia lysates from ectocervical organoids, with or without pre-treatment with human β-defensins (HBD1, HBD2, or both) at 5 days post-infection (dpi). Data represent mean ± s.d. from 8 non-overlapping regions across two replicates. f) Violin plot showing gene set enrichment scores for the GO term related to the adaptive immune response across epithelial subclusters under different infection conditions. g) Trend plot showing changes in mean expression levels of selected proliferation markers MKI67 and CDK1 under different infection conditions; Lines are colored by gene; point shapes distinguish ectocervical (circles) and endocervical (triangles) samples.

Article Snippet: The following primary antibodies were used for immunofluorescence: mouse anti-acetylated tubulin-Alexa-647 (Santa Cruz Biotechnology, sc-23950-AF647), mouse anti-E-cadherin-Alexa-488 (BD Biosciences, 560061), mouse anti-E-cadherin (BD Biosciences, 610181), rabbit anti-KRT5-Alexa488 (Abcam, ab193894 mouse anti-MUC5B (Abcam, ab77995), rabbit anti-MUC21 (ProteinAtlas, HPA052028), rabbit anti-KRT8 (1:200, Abcam, abab59400), mouse-anti-KRT6 (Abcam, ab18586), recombinant rabbit anti-PAX8 (Abcam, ab239363), goat anti- Chlamydia trachomatis Major Outer Membrane Protein (MOMP) (1:500, BIO-RAD, 1990-0804) and for labeling the DNA, DAPI (Roche, 10236276001) was used.

Techniques: Expressing, Quantitative RT-PCR, Infection, Control

Journal: bioRxiv

Article Title: Single-cell atlas of patient-derived cervical organoids uncovers epithelial immune heterogeneity and intercellular crosstalk during Chlamydia infection

doi: 10.1101/2025.04.13.648603

Figure Lengend Snippet: a-b) Chord diagrams showing upregulated (a) and downregulated (b) ligand-receptor (L-R) signaling pairs in ectocervical squamous epithelial cells upon Chlamydia infection; Outer bars indicate signal sending (ligand-expressing) cell groups; inner bars colored by receiving (receptor-expressing) cell groups; edges colored by signaling source/senders. c-d) River plots depicting the inferred outgoing (c) and incoming (d) communication patterns in ectocervical squamous epithelia, highlighting the associations between latent patterns, cell groups, and enriched signaling pathways; thickness of the flow reflects the contribution of cell groups or signaling pathways to each pattern, with pathway labels colored by signaling category. e) Heatmap showing the relative importance and signaling role of each cell group in the MIF, SEMA6, and CADM signaling networks in the ectocervix; scale bar color denotes the criticality of a cell group in driving the communication network ranging from high (dark green) to low (gray). f-g) Chord diagrams illustrating upregulated (f) and downregulated (g) L–R signaling interactions across endocervical columnar epithelial subpopulations after infection. h) Scatter plot visualization of predominant signal senders and receivers among endocervical epithelial subsets in 2D space; Circle size represents the total inferred links (outgoing and incoming) associated with each cell type, colored by subclusters. i-j) Inferred outgoing (i) and incoming (j) communication patterns in endocervical epithelia, illustrating the links between latent patterns, cell types, and signaling pathways. k) Heatmap visualization of the relative importance and signaling role of each cell group in the endocervix for IL-1, MIF, and TNF signaling networks. Scale bar color as in (e).

Article Snippet: The following primary antibodies were used for immunofluorescence: mouse anti-acetylated tubulin-Alexa-647 (Santa Cruz Biotechnology, sc-23950-AF647), mouse anti-E-cadherin-Alexa-488 (BD Biosciences, 560061), mouse anti-E-cadherin (BD Biosciences, 610181), rabbit anti-KRT5-Alexa488 (Abcam, ab193894 mouse anti-MUC5B (Abcam, ab77995), rabbit anti-MUC21 (ProteinAtlas, HPA052028), rabbit anti-KRT8 (1:200, Abcam, abab59400), mouse-anti-KRT6 (Abcam, ab18586), recombinant rabbit anti-PAX8 (Abcam, ab239363), goat anti- Chlamydia trachomatis Major Outer Membrane Protein (MOMP) (1:500, BIO-RAD, 1990-0804) and for labeling the DNA, DAPI (Roche, 10236276001) was used.

Techniques: Infection, Expressing, Protein-Protein interactions

Journal: bioRxiv

Article Title: Single-cell atlas of patient-derived cervical organoids uncovers epithelial immune heterogeneity and intercellular crosstalk during Chlamydia infection

doi: 10.1101/2025.04.13.648603

Figure Lengend Snippet: (a) Anatomical overview of the human uterine cervix highlighting the ectocervix and endocervix, their distinct epithelial compositions, and convergence at the transition zone. Patient-derived 3D organoids derived from adult epithelial stem cells preserve region-specific epithelial architecture and identity. (b) Integration of organoid and cervical tissue scRNA-seq datasets reveals high transcriptional fidelity and molecular congruence across corresponding epithelial subsets. Gene signature and similarity analyses demonstrate that organoids recapitulate both cellular heterogeneity and native epithelial programs. (c) Depiction shows the identified lineage-specific epithelial subtypes across squamous and columnar compartments under homeostasis. Pseudotime trajectory analysis reveals hierarchical differentiation from basal/stem-like cells to terminally differentiated states, with key subtype-defining transcriptional markers shown. (d) Depiction highlighting region- and cell-type-resolved expression profiles of innate immune defense genes at steady state. Distinct epithelial subtypes show selective enrichment of mucins, junctional components, PRRs, antimicrobial peptides (AMPs), and cytokines, reflecting compartmentalized mucosal immune strategies. (e) Schematic showing that Chlamydia infection remodels the epithelial landscape. In the endocervix, transcriptional reprogramming of uninfected bystander cells gives rise to IFN-responsive columnar subsets with robust ISG expression, hallmarks of paracrine immune activation. (f) Infection reconfigures epithelial communication networks. Cell-cell interaction modeling reveals that columnar bystander cells function as central signaling hubs mediating immune and regenerative responses through pathways such as CCL, CXCL, MHC-II, MIF, SEMA, TNF and IL-1. Region-specific shifts in ligand-receptor interactions underscore divergent mucosal responses between ecto- and endocervix.

Article Snippet: The following primary antibodies were used for immunofluorescence: mouse anti-acetylated tubulin-Alexa-647 (Santa Cruz Biotechnology, sc-23950-AF647), mouse anti-E-cadherin-Alexa-488 (BD Biosciences, 560061), mouse anti-E-cadherin (BD Biosciences, 610181), rabbit anti-KRT5-Alexa488 (Abcam, ab193894 mouse anti-MUC5B (Abcam, ab77995), rabbit anti-MUC21 (ProteinAtlas, HPA052028), rabbit anti-KRT8 (1:200, Abcam, abab59400), mouse-anti-KRT6 (Abcam, ab18586), recombinant rabbit anti-PAX8 (Abcam, ab239363), goat anti- Chlamydia trachomatis Major Outer Membrane Protein (MOMP) (1:500, BIO-RAD, 1990-0804) and for labeling the DNA, DAPI (Roche, 10236276001) was used.

Techniques: Derivative Assay, Expressing, Infection, Activation Assay