Journal: Animals : an Open Access Journal from MDPI

Article Title: Induction of Serum Amyloid A3 in Mouse Mammary Epithelial Cells Stimulated with Lipopolysaccharide and Lipoteichoic Acid

doi: 10.3390/ani11061548

Figure Lengend Snippet: Alignment of amino acid sequences of mouse serum amyloid A (SAA) isoforms. The sequences (AAEKISDGREAFOE and QRWVQFMKEAG) of synthetic SAA1 and SAA3 peptides for immunization to rabbits are underlined. Accession numbers are shown in parentheses.

Article Snippet: Subsequently, the membrane was incubated with the primary anti-mouse SAA1 rabbit polyclonal antibody (1:500, #PAA885Mu01, Cloud-Clone, Houston, TX, USA) or anti-SAA3 rat monoclonal antibody [JOR110A] (1:50, #ab231680, Abcam, Cambridge, UK) diluted with 1% nonfat dried skim milk in TBST for 1 h at room temperature.

Techniques:

Journal: Animals : an Open Access Journal from MDPI

Article Title: Induction of Serum Amyloid A3 in Mouse Mammary Epithelial Cells Stimulated with Lipopolysaccharide and Lipoteichoic Acid

doi: 10.3390/ani11061548

Figure Lengend Snippet: Oligonucleotide primers used in quantitative real-time PCR.

Article Snippet: Subsequently, the membrane was incubated with the primary anti-mouse SAA1 rabbit polyclonal antibody (1:500, #PAA885Mu01, Cloud-Clone, Houston, TX, USA) or anti-SAA3 rat monoclonal antibody [JOR110A] (1:50, #ab231680, Abcam, Cambridge, UK) diluted with 1% nonfat dried skim milk in TBST for 1 h at room temperature.

Techniques: Sequencing

Journal: Animals : an Open Access Journal from MDPI

Article Title: Induction of Serum Amyloid A3 in Mouse Mammary Epithelial Cells Stimulated with Lipopolysaccharide and Lipoteichoic Acid

doi: 10.3390/ani11061548

Figure Lengend Snippet: Comparison of Saa1 and Saa3 mRNA expression levels in NMuMG cells treated with lipopolysaccharide (LPS) and lipoteichoic acid (LTA). Data are presented as the mean ± standard deviation of four independent experiments. Asterisks indicate significant difference compared with control levels: ** p < 0.01. NMuMG, normal murine mammary gland.

Article Snippet: Subsequently, the membrane was incubated with the primary anti-mouse SAA1 rabbit polyclonal antibody (1:500, #PAA885Mu01, Cloud-Clone, Houston, TX, USA) or anti-SAA3 rat monoclonal antibody [JOR110A] (1:50, #ab231680, Abcam, Cambridge, UK) diluted with 1% nonfat dried skim milk in TBST for 1 h at room temperature.

Techniques: Expressing, Standard Deviation

Journal: Animals : an Open Access Journal from MDPI

Article Title: Induction of Serum Amyloid A3 in Mouse Mammary Epithelial Cells Stimulated with Lipopolysaccharide and Lipoteichoic Acid

doi: 10.3390/ani11061548

Figure Lengend Snippet: Comparison of SAA1 (A and B) and SAA3 (C and D) protein expression levels in NMuMG cells treated with lipopolysaccharide (LPS) or lipoteichoic acid (LTA) by immunofluorescence analysis (IFA). ( A , C ) Fields of view where fluorescence intensity was measured. Scale bar = 20 µm. ( B , D ) The relative SAA1 and SAA3 protein expression levels in NMuMG cells following stimulation with LPS or LTA were normalized to those in untreated, control cells. Data are presented as the mean fluorescence of five or more random locations with vertical bars representing standard deviation. Asterisks indicate significant difference compared with control levels: * p < 0.05, ** p < 0.01. NMuMG, normal murine mammary gland.

Article Snippet: Subsequently, the membrane was incubated with the primary anti-mouse SAA1 rabbit polyclonal antibody (1:500, #PAA885Mu01, Cloud-Clone, Houston, TX, USA) or anti-SAA3 rat monoclonal antibody [JOR110A] (1:50, #ab231680, Abcam, Cambridge, UK) diluted with 1% nonfat dried skim milk in TBST for 1 h at room temperature.

Techniques: Expressing, Immunofluorescence, Fluorescence, Standard Deviation

Journal: Animals : an Open Access Journal from MDPI

Article Title: Induction of Serum Amyloid A3 in Mouse Mammary Epithelial Cells Stimulated with Lipopolysaccharide and Lipoteichoic Acid

doi: 10.3390/ani11061548

Figure Lengend Snippet: Detection of SAA1 and SAA3 protein expression in NMuMG cell supernatants treated with lipopolysaccharide (LPS) or lipoteichoic acid (LTA) by Western blot. ( A ) Detection of SAA1 in the cell supernatant. Membranes were incubated with the primary anti-mouse SAA1 antibody (dilution of 1:500, 60 s exposure time). ( B ) Detection of SAA3 in the cell supernatant. Membranes were incubated with the primary anti-mouse SAA3 antibody (dilution of 1:50, 120 s exposure time). PC, positive control; 1, recombinant murine SAA1 (rSAA1); 3, rSAA3; NMuMG, normal murine mammary gland.

Article Snippet: Subsequently, the membrane was incubated with the primary anti-mouse SAA1 rabbit polyclonal antibody (1:500, #PAA885Mu01, Cloud-Clone, Houston, TX, USA) or anti-SAA3 rat monoclonal antibody [JOR110A] (1:50, #ab231680, Abcam, Cambridge, UK) diluted with 1% nonfat dried skim milk in TBST for 1 h at room temperature.

Techniques: Expressing, Western Blot, Incubation, Positive Control, Recombinant

Journal: iScience

Article Title: Single-cell RNA sequencing reveals epithelial cells driving brain metastasis in lung adenocarcinoma

doi: 10.1016/j.isci.2024.109258

Figure Lengend Snippet: Driver genes of BMAECs analysis Metastasis-associated ECs (A) reclustering analysis and (B) pseudotime trajectory analysis. (C) KEGG enrichment analysis of BMAECs. (D) Specific gene display of BMAECs. SPP1 (E) SAA1 (F) and CDKN2A (G) expression intensity in BMAECs.

Article Snippet: Immunohistochemistry assay was performed on 10% formalin fixed, paraffin-embedded (FFPE) sections (26 pLUADs and 20 pLUADs samples in which BM occurred)., The antibodies used in each samples were: anti-SPP1 (22952-1-AP, Proteintech, 1:200), anti-SAA1 (R381812, Zenbio, 1:200), anti-CDKN2A (10883-1-AP, Proteintech, 1:500) and secondary antibody.

Techniques: Expressing

Journal: iScience

Article Title: Single-cell RNA sequencing reveals epithelial cells driving brain metastasis in lung adenocarcinoma

doi: 10.1016/j.isci.2024.109258

Figure Lengend Snippet: Validation of BM driver genes in LUAD associated with metastasis and prognosis (A) Representative IHC staining plots of SPP1, SAA1, and CDKN2A in samples of pLUAD (left panel), pLUAD of BM (right panel). Cell nuclei were counterstained with DAPI (blue). Scale bar: 5 μm. (B) SPP1 (C) SAA1 and (D) CDKN2A H-scores are represented by bar graphs. ∗ indicates p < 0.05, ∗∗ indicates p < 0.01, and ∗∗∗ indicates p < 0.001. BM cumulative probability curves are shown in (E) (F) and (G) plots (log rank p < 0.0001, p = 0.0230, p = 0.0180, respectively). (H) Representative IF confocal staining plots for markers SPP1 (red), SAA1 (green), CDKN2A (pink) in pLUAD (top) and BM (bottom) samples. Cell nuclei were counterstained with DAPI (blue). Scale bar: 80 μm (left panel), 20 μm (right panel).

Article Snippet: Immunohistochemistry assay was performed on 10% formalin fixed, paraffin-embedded (FFPE) sections (26 pLUADs and 20 pLUADs samples in which BM occurred)., The antibodies used in each samples were: anti-SPP1 (22952-1-AP, Proteintech, 1:200), anti-SAA1 (R381812, Zenbio, 1:200), anti-CDKN2A (10883-1-AP, Proteintech, 1:500) and secondary antibody.

Techniques: Immunohistochemistry, Staining

Journal: iScience

Article Title: Single-cell RNA sequencing reveals epithelial cells driving brain metastasis in lung adenocarcinoma

doi: 10.1016/j.isci.2024.109258

Figure Lengend Snippet:

Article Snippet: Immunohistochemistry assay was performed on 10% formalin fixed, paraffin-embedded (FFPE) sections (26 pLUADs and 20 pLUADs samples in which BM occurred)., The antibodies used in each samples were: anti-SPP1 (22952-1-AP, Proteintech, 1:200), anti-SAA1 (R381812, Zenbio, 1:200), anti-CDKN2A (10883-1-AP, Proteintech, 1:500) and secondary antibody.

Techniques: Paraffin Section, RNA Sequencing Assay, Software