Journal: Cell death discovery

Article Title: Inhibition of Aurora Kinase B activity disrupts development and differentiation of salivary glands.

doi: 10.1038/s41420-020-00393-w

Figure Lengend Snippet: Fig. 8 AURKB loss disrupted acinar identity via ROS generation. A, B Mist1 (green) and pan cytokeratin (blue) expressions in E16.5+24h bud cells, scale bar = 50 µm. Remarkable loss of the acinar differentiation marker Mist1 at the protein and transcriptional levels in response to Barasertib treatment. Recovery of Mist1 levels due to ROS scavenging using aminoguanidine (number of pooled glands ≥5), analysed by ANOVA **P ≤0.01, *p ≤0.05, ****P ≤0.0001. C, D ROS production in response to AURKB inhibition in the embryonic salivary glands. ROS levels were visualised in the SMG explants after staining with DCFDA and inspection under fluorescence microscope. Analysis of corrected total fluorescence (CTF) using ImageJ revealed that Barasertib treatment for 24 h significantly increased levels of ROS in the E16.5+24h explants which were significantly reduced after co- treating the explants with aminoguanidine, analysed by ANOVA, **P ≤0.01, *p ≤0.05, ns: non-significant, p > 0.05. Scale bar = 500 μm, n = 3/group.

Article Snippet: To block endogenous peroxidase activity and nonspecific background staining sections were incubated in 3% hydrogen peroxide solution for 20–30 min. To block all epitopes on the tissue samples and prevent nonspecific antibody binding, sections were incubated with 1% BSA in 1X TBS, pH 7.6 for 5 min. Tissue sections were incubated at 4 °C overnight, with the antibody: rabbit-anti-cleaved caspase 3 (1:2500, NB100–56113, Novus Bio), rabbit–anti-Mist1 (1:200, ab187978, Abcam), mouse-anti-Cyclin D1 (1:400, NBP2–32840, Novus Bio), rabbit-anti-AURKB (1:200, ab115793, Abcam), rabbit-anti-ki67 (1:100, ab16667, Abcam), rabbit-anti-p21 (1:200, ab188224, Abcam), rabbit-anti-p.H2AX(1:200, ab81299, Abcam), rat-anti-E- Cadherin (1:200, sc-59778, Santa Cruz Biotechnology, Inc.) guinea pig-anti-pan cytokeratin (1:50, BP5069, Origene).

Techniques: Marker, Inhibition, Staining, Microscopy

Journal: International journal of oncology

Article Title: Oncolytic herpes simplex virus treatment of metastatic breast cancer.

doi: 10.3892/ijo.2011.1266

Figure Lengend Snippet: Figure 1. Establishment of HMEC culture. (A) Primary culture of HMEC1 (the arrow shows edge of biopsy). (B) Morphology of HMEC1 (passage 2). (C) Positive staining of cytokeratin in HMEC1 cytoplasm with the nucleus stained by hematoxylin. (D) Morphology of cells undergoing senescence and death.

Article Snippet: The cells that grew out from the biopsies were named HMEC1 and HMEC2 and characterized with a mouse anti-cytokeratin antibody (Zhongshan Goldenbridge Biotechnology, Beijing, AE1/AE3, ZM-0069).

Techniques: Staining

Journal: bioRxiv

Article Title: Spatial transcriptomic analysis of progressing oral epithelial dysplasia reveals unique differentially expressed genes and microenvironmental changes

doi: 10.64898/2026.01.07.697832

Figure Lengend Snippet: Differential gene expression in progressors (biopsy 1) and non-progressors (biopsy 1) within PanCK-positive (A-C) and PanCK-negative (D) regions of interest (ROI). (A) Heatmap depicting unsupervised clustering of samples into two groups based on differentially expressed (DE) genes. (B) Volcano plot highlighting significantly DE genes with yellow dots. (C) Violin plots illustrating the most prominently downregulated gene (CDKN2A) and upregulated gene (STOM) in progressing oral epithelial dysplasia (OED). (D) Volcano plot displaying significantly DE genes with yellow dots, alongside violin plots showing downregulated genes (S100A10 and KL4) and upregulated gene (ZFAT) in progressing OED.

Article Snippet: Slide preparation was performed according to the manufacturer’s protocol (Nanostring GeoMx) with the following digestion and heat-induced epitope retrieval (HIER) conditions: 0.1 μg/ml Proteinase K for 15 min and ER for 20 min. ROI selection (PanCK+/PanCK-) was performed in the GeoMx Digital Spatial Profiler (DSP) based on morphology and immunofluorescence staining with PanCK (Novus Biologicals, clone AE1/AE3, NBP2-33200AF488), SMA (Abcam, clone 1A4, ab202368), CD45 (CST, clone D9M8I, 13917BF), and SYTO 83 DNA staining (ThermoFisher, S11364).

Techniques: Gene Expression

Journal: bioRxiv

Article Title: Spatial transcriptomic analysis of progressing oral epithelial dysplasia reveals unique differentially expressed genes and microenvironmental changes

doi: 10.64898/2026.01.07.697832

Figure Lengend Snippet: (A) Bar graph showing the most enriched pathways in Progressors, indicating the number of enriched pathways within each category. (B) Heatmap illustrating clustering of PanCK-positive (PanCK+) regions of interest (ROI) in progressing and non-progressing oral epithelial dysplasia (OED). (C) Volcano plot displaying differentially expressed (DE) pathways, with yellow dots representing significant pathways in PanCK+ ROI of progressors and non-progressors. (D) Heatmap showing clustering of PanCK-negative (PanCK-) ROI in progressing and non-progressing OED. (E) Heatmap illustrating clustering of PanCK+ ROI in precursor lesions and subsequent oral squamous cell carcinoma (OSCC) samples. (F) Heatmap depicting clustering of PanCK- ROI in precursor lesions and subsequent OSCC samples. (G) Volcano plot with yellow dots indicating DE pathways in PanCK+ ROI of precursor lesions and subsequent OSCC.

Article Snippet: Slide preparation was performed according to the manufacturer’s protocol (Nanostring GeoMx) with the following digestion and heat-induced epitope retrieval (HIER) conditions: 0.1 μg/ml Proteinase K for 15 min and ER for 20 min. ROI selection (PanCK+/PanCK-) was performed in the GeoMx Digital Spatial Profiler (DSP) based on morphology and immunofluorescence staining with PanCK (Novus Biologicals, clone AE1/AE3, NBP2-33200AF488), SMA (Abcam, clone 1A4, ab202368), CD45 (CST, clone D9M8I, 13917BF), and SYTO 83 DNA staining (ThermoFisher, S11364).

Techniques:

Journal: Cancer Research

Article Title: Tissue Factor Regulation by Epidermal Growth Factor Receptor and Epithelial-to-Mesenchymal Transitions: Effect on Tumor Initiation and Angiogenesis

doi: 10.1158/0008-5472.can-08-2067

Figure Lengend Snippet: Figure 3. Changes in TF expression associated with multilineage differentiation of cancer cells in vivo. A, the majority of cells within the tumor mass were human cancer cells, as determined by staining with an antibody directed against human (tumor) cell nuclear antigen (green). Tumors grown in YFP/SCID mice were double-stained using anti-TF (human cancer cells, green) and anti-GFP (host, red) antibodies. Infiltration of YFP-positive host cells in A431 tumors is minimal (<10%) and many stroma-poor tumor regions remain negative for TF. Immunofluorescence staining of cytokeratin (red) reveals highly keratinized pearl structures within these tumors. B, some tumor cells have lost their epithelial phenotype and express the mesenchymal marker vimentin (green), indicative of cells undergoing EMT. H&E staining depicts which regions of the tumor are likely to express vimentin and undergo EMT (tumor periphery) and those that retain their epithelial phenotype and differentiate along an epithelial pathway (central tumor). Arrowhead, a keratin pearl. Cytokeratin immunofluorescence (red) shows regions of the tumor that have undergone epithelial differentiation. C, comparative Western blot analysis between A431 cells maintained in culture and those grown as tumors. A global decrease in TF protein levels were observed in tumors. Changes in EGFR were unremarkable whereas the expression of E-cadherin is reduced and the extracellular region is cleaved, leaving an 80 kDa protein that is unable to make cell-cell contacts. Consistent with this loss of E-cadherin and epithelial phenotype, some tumor regions express vimentin, whereas other regions of these tumors remain epithelial and express high levels of cytokeratin. Bars, 100 AM.

Article Snippet: All other tissue staining was performed on 4-Am paraffin sections which were incubated overnight at 4jC with the respective primary antibodies following heat antigen retrieval; except for cytokeratin (Novus Biological, Inc.) staining which required proteolytic digestion prior to a 60 min incubation with the prediluted antibody at room temperature.

Techniques: Expressing, In Vivo, Staining, Immunofluorescence, Marker, Western Blot