Journal: Oncology Letters

Article Title: ANGPTL4 overexpression is associated with progression and poor prognosis in breast cancer

doi: 10.3892/ol.2020.11768

Figure Lengend Snippet: ANGPTL4 expression in normal breast (n=40), ADH (n=40), DCIS (n=40) and IDC (n=205) tissues.

Article Snippet: Non-specific binding sites were blocked by incubation with 10% normal goat serum (cat. no. ab7481; Abcam) at room temperature for 20 min. Tissue sections were then incubated with a primary ANGPTL4 polyclonal antibody (dilution, 1:100; cat. no. A01147; Boster Biological Technology) at 37°C for 2 h. All sections were subsequently incubated using the Dako REALTM EnVisionTM Detection System (cat. no. K5007; Dako; Agilent Technologies, Inc.) containing biotinylated anti-rabbit secondary antibodies (dilution, 1:100) for 20 min at 37°C, and peroxidase-labelled streptavidin for 20 min at room temperature (dilution, 1:100).

Techniques: Expressing

Journal: Oncology Letters

Article Title: ANGPTL4 overexpression is associated with progression and poor prognosis in breast cancer

doi: 10.3892/ol.2020.11768

Figure Lengend Snippet: The IHC staining of angiopoietin-like 4 expression in normal breast, atypical ductal hyperplasia and ductal carcinoma in situ tissues. Angiopoietin-like 4 in normal breast tissues: (A1) Negative, (A2) positive; in atypical ductal hyperplasia tissues: (B1) Negative, (B2) positive; and in ductal carcinoma in situ tissues: (C1) Negative, (C2) positive. Magnification, x200.

Article Snippet: Non-specific binding sites were blocked by incubation with 10% normal goat serum (cat. no. ab7481; Abcam) at room temperature for 20 min. Tissue sections were then incubated with a primary ANGPTL4 polyclonal antibody (dilution, 1:100; cat. no. A01147; Boster Biological Technology) at 37°C for 2 h. All sections were subsequently incubated using the Dako REALTM EnVisionTM Detection System (cat. no. K5007; Dako; Agilent Technologies, Inc.) containing biotinylated anti-rabbit secondary antibodies (dilution, 1:100) for 20 min at 37°C, and peroxidase-labelled streptavidin for 20 min at room temperature (dilution, 1:100).

Techniques: Immunohistochemistry, Expressing, In Situ

Journal: Oncology Letters

Article Title: ANGPTL4 overexpression is associated with progression and poor prognosis in breast cancer

doi: 10.3892/ol.2020.11768

Figure Lengend Snippet: Angiopoietin-like 4 expression in invasive ductal carcinoma. (A1) Negative (magnification, x200); (A2) negative (magnification, x400); (B1) weakly positive (magnification, x200); (B2) weakly positive (magnification, x400); (C1) moderately positive (magnification, x200); (C2) moderately positive (magnification, x400); (D1) strongly positive (magnification, x200); (D2) strongly positive (magnification, x400).

Article Snippet: Non-specific binding sites were blocked by incubation with 10% normal goat serum (cat. no. ab7481; Abcam) at room temperature for 20 min. Tissue sections were then incubated with a primary ANGPTL4 polyclonal antibody (dilution, 1:100; cat. no. A01147; Boster Biological Technology) at 37°C for 2 h. All sections were subsequently incubated using the Dako REALTM EnVisionTM Detection System (cat. no. K5007; Dako; Agilent Technologies, Inc.) containing biotinylated anti-rabbit secondary antibodies (dilution, 1:100) for 20 min at 37°C, and peroxidase-labelled streptavidin for 20 min at room temperature (dilution, 1:100).

Techniques: Expressing

Journal: Oncology Letters

Article Title: ANGPTL4 overexpression is associated with progression and poor prognosis in breast cancer

doi: 10.3892/ol.2020.11768

Figure Lengend Snippet: Association between ANGPTL4 expression and clinical parameters in 205 patients with invasive ductal carcinoma.

Article Snippet: Non-specific binding sites were blocked by incubation with 10% normal goat serum (cat. no. ab7481; Abcam) at room temperature for 20 min. Tissue sections were then incubated with a primary ANGPTL4 polyclonal antibody (dilution, 1:100; cat. no. A01147; Boster Biological Technology) at 37°C for 2 h. All sections were subsequently incubated using the Dako REALTM EnVisionTM Detection System (cat. no. K5007; Dako; Agilent Technologies, Inc.) containing biotinylated anti-rabbit secondary antibodies (dilution, 1:100) for 20 min at 37°C, and peroxidase-labelled streptavidin for 20 min at room temperature (dilution, 1:100).

Techniques: Expressing

Journal: Oncology Letters

Article Title: ANGPTL4 overexpression is associated with progression and poor prognosis in breast cancer

doi: 10.3892/ol.2020.11768

Figure Lengend Snippet: Survival curves based on ANGPTL4 expression. (A) DFS and (B) OS analysis of ANGPTL4 in breast cancer. ANGPTL4, angiopoietin-like 4; DFS, disease-free survival; OS, overall survival.

Article Snippet: Non-specific binding sites were blocked by incubation with 10% normal goat serum (cat. no. ab7481; Abcam) at room temperature for 20 min. Tissue sections were then incubated with a primary ANGPTL4 polyclonal antibody (dilution, 1:100; cat. no. A01147; Boster Biological Technology) at 37°C for 2 h. All sections were subsequently incubated using the Dako REALTM EnVisionTM Detection System (cat. no. K5007; Dako; Agilent Technologies, Inc.) containing biotinylated anti-rabbit secondary antibodies (dilution, 1:100) for 20 min at 37°C, and peroxidase-labelled streptavidin for 20 min at room temperature (dilution, 1:100).

Techniques: Expressing

Journal: Oncology Letters

Article Title: ANGPTL4 overexpression is associated with progression and poor prognosis in breast cancer

doi: 10.3892/ol.2020.11768

Figure Lengend Snippet: Cox proportional hazards regression analysis for DFS and OS.

Article Snippet: Non-specific binding sites were blocked by incubation with 10% normal goat serum (cat. no. ab7481; Abcam) at room temperature for 20 min. Tissue sections were then incubated with a primary ANGPTL4 polyclonal antibody (dilution, 1:100; cat. no. A01147; Boster Biological Technology) at 37°C for 2 h. All sections were subsequently incubated using the Dako REALTM EnVisionTM Detection System (cat. no. K5007; Dako; Agilent Technologies, Inc.) containing biotinylated anti-rabbit secondary antibodies (dilution, 1:100) for 20 min at 37°C, and peroxidase-labelled streptavidin for 20 min at room temperature (dilution, 1:100).

Techniques:

Journal: Cell Discovery

Article Title: Luminal hormone-responsive cells tune the regenerative remodeling of mammary glands in large mammals

doi: 10.1038/s41421-025-00848-3

Figure Lengend Snippet: a UMAP plot displaying 14 identified cell types within the goat mammary glands. Cells are annotated and colored by type. Subsets of cell types including luminal, basal, fibroblast, immune, endothelial cell are labeled. b The percentages of −4W (red) and +1 W (blue) cells in each cell type are shown in a UMAP plot. c UMAP plots showing the expression of selected marker genes in four luminal subtypes. d Pseudotemporal trajectory analysis of scRNA-seq data of luminal secretory cells is shown in a UMAP plot. e Changes in the proportion of LumSecP and LumSec cells in luminal cell types were identified by scRNA-seq data at −4W and +1 W. n = 3 goats per group. f Representative images of tissue immunofluorescence staining for ALDH1A3 (red), KRT18 (green), and DAPI at −4W and +1 W. Scale bars, 50 μm. g Bar plots exhibiting the percentage of ALDH1A3-positive cells in luminal cells (labeled by KRT18) in f . n = 15 sections per group. h Representative images of tissue immunofluorescence staining for FABP3 (red), KRT18 (green), and DAPI at −4W and +1 W. Scale bars, 50 μm. i Bar plots exhibiting the percentage of FABP3-positive cells in luminal cells (labeled by KRT18) in h . n = 20 sections per group. The data are presented as the mean ± SEM. The P values of two-sided Student’s t -tests are shown in e , g , i .

Article Snippet: For immunofluorescence staining, goat mammary tissues were stained with primary antibodies against ALDH1A3 (#25167-1-AP, Proteintech), FABP3 (#10676-1-AP, Proteintech), KRT17 (#17516-1-AP, Proteintech), KRT14 (#SC65-06, HUABIO), KRT18 (conjugated with iFluorTM 488, #SZ80-07, HUABIO), KRT8 (#10384-1-AP, ProteinTech), PR (#25871-1-AP, ProteinTech), ER (#21244-1-AP, ProteinTech), and PCNA (#ab29, Abcam).

Techniques: Labeling, Expressing, Marker, Immunofluorescence, Staining

Journal: Cell Discovery

Article Title: Luminal hormone-responsive cells tune the regenerative remodeling of mammary glands in large mammals

doi: 10.1038/s41421-025-00848-3

Figure Lengend Snippet: a Relative proportions of LumHR cells in total luminal cells identified by scRNA-seq data at −4W and +1 W. n = 3 goats per group. b , c Bar plots exhibiting the percentage of PGR - and ESR1 -positive cells within luminal cells in scRNA-seq data. n = 3 goats per group. d Representative images of immunofluorescence staining for PR (red), KRT18 (green), and DAPI (blue). Scale bars, 50 μm. e Bar plots exhibiting the percentage of PR-positive cells in luminal cells (labeled by KRT18) in ( d ). n = 5 goats per group. f Representative images of immunofluorescence staining for ER (red), KRT18 (green), and DAPI (blue). Scale bars, 50 μm. g Bar plots exhibiting the percentage of ER-positive cells in luminal cells (labeled by KRT18) in ( f ). n = 5 goats. h Violin plot showing the specific expression of PRLR in LumHR cells by scRNA-seq. i Heatmap displaying the transcriptional level of indicated genes related to milk protein and luminal differentiation in the goat mammary organoids ( n = 3 biological replicates) treated with or without prolactin and in the mammary tissue at −4W (non-lactation) and +1 W (lactation). n = 3 goats for tissues. j Proportions of luminal cell types in goat mammary organoids incubated with or without prolactin predicted by CIBERSORTx deconvolution. n = 3 biological replicates. k Representative images of immunofluorescence staining for ALDH1A3 (red), KRT18 (green), and DAPI in mammary organoids incubated with or without prolactin. Scale bars, 10 μm. l Bar plots exhibiting the percentage of ALDH1A3-positive cells in luminal cells (labeled by KRT18) in ( k ). n = 14 domes in the control group and n = 10 domes in the prolactin treated group. m Representative images of immunofluorescence staining for FABP3 (red), KRT18 (green) and DAPI in mammary organoids incubated with or without prolactin. Scale bars, 10 μm. n Bar plots exhibiting the percentage of FABP3-positive cells in luminal cells (labeled by KRT18) in ( m ). n = 6 domes in the control group and n = 7 domes in the prolactin-treated group. The data are presented as the mean ± SEM. The P values of two-sided Student’s t -tests are shown in ( a – c , e , g , j , l , n ).

Article Snippet: For immunofluorescence staining, goat mammary tissues were stained with primary antibodies against ALDH1A3 (#25167-1-AP, Proteintech), FABP3 (#10676-1-AP, Proteintech), KRT17 (#17516-1-AP, Proteintech), KRT14 (#SC65-06, HUABIO), KRT18 (conjugated with iFluorTM 488, #SZ80-07, HUABIO), KRT8 (#10384-1-AP, ProteinTech), PR (#25871-1-AP, ProteinTech), ER (#21244-1-AP, ProteinTech), and PCNA (#ab29, Abcam).

Techniques: Immunofluorescence, Staining, Labeling, Expressing, Incubation, Control

Journal: Cell Discovery

Article Title: Luminal hormone-responsive cells tune the regenerative remodeling of mammary glands in large mammals

doi: 10.1038/s41421-025-00848-3

Figure Lengend Snippet: a Schematic illustration of targeted ablation of LumHR cells using the prlr -promoter to drive expression of DTA. b Experimental setup used in AAV intraductally injected mammary gland of ROSA-DTA +/ − mice under RR. c Whole-mount staining with carmine alum of mammary glands from ROSA-DTA +/ − mice (lactation day 2) intraductally injected with AAV-pPrlr-Cre or AAV-Control . Scale bars, 4 mm (top) and 500 μm (bottom). d , e Immunohistochemical staining and quantification of ER-positive luminal cells in mammary glands intraductally injected with AAV-pPrlr-Cre or AAV-Control. n = 3 mice per group. Scale bars, 50 μm. f , g Immunohistochemical staining and quantification of PR-positive luminal cells in mammary glands intraductally injected with AAV-pPrlr-Cre or AAV-Control . n = 3 mice per group. Scale bars, 50 μm. h , i Immunohistochemical staining and quantification of β-casein-positive alveoli number per mm 2 in mammary glands intraductally injected with AAV-pPrlr-Cre or AAV-Control . n = 3 mice per group. Scale bars, 50 μm. j Representative images of immunofluorescence staining for FABP3 (red), KRT18 (green), and DAPI in mammary glands intraductally injected with AAV-pPrlr-Cre or AAV-Control . Scale bars, 50 μm. k Bar plots exhibiting the percentage of FABP3-positive cells in luminal cells (labeled by KRT18) in j . n = 3 mice per group. l Representative images of immunofluorescence staining for ALDH1A3 (red), KRT18 (green), and DAPI in mammary glands intraductally injected with AAV-pPrlr-Cre or AAV-Contro l. Scale bars, 50 μm. m Bar plots exhibiting the percentage of FABP3-positive cells in luminal cells (labeled by KRT18) in ( l ). n = 3 mice per group. The data are presented as the mean ± SEM. The P values of two-sided Student’s t -tests are shown in d , g , i , l , m .

Article Snippet: For immunofluorescence staining, goat mammary tissues were stained with primary antibodies against ALDH1A3 (#25167-1-AP, Proteintech), FABP3 (#10676-1-AP, Proteintech), KRT17 (#17516-1-AP, Proteintech), KRT14 (#SC65-06, HUABIO), KRT18 (conjugated with iFluorTM 488, #SZ80-07, HUABIO), KRT8 (#10384-1-AP, ProteinTech), PR (#25871-1-AP, ProteinTech), ER (#21244-1-AP, ProteinTech), and PCNA (#ab29, Abcam).

Techniques: Expressing, Injection, Staining, Control, Immunohistochemical staining, Immunofluorescence, Labeling