mouse brca1 cdna  (OriGene)

Journal: Nature communications

Article Title: S100A9-CXCL12 activation in BRCA1-mutant breast cancer promotes an immunosuppressive microenvironment associated with resistance to immunotherapy.

doi: 10.1038/s41467-022-29151-5

Figure Lengend Snippet: Fig. 3 S100a9 gene is regulated by both Brca1 and S100a9. a Expression of S100a9 and S100a8 in Brca1 MT (G600) and WT (B477) mammary epithelial cell lines. b Expression of S100a9 and S100a8 in B477 cells with the expression of shBrca1 at different concentrations. c Expression of S100a9 and S100a8 in B477 cells in which the mBrca1 gene was overexpressed. d Expression of S100A9 and S100A8 in MDA-MB-231 (231) control and 231 cells with the expression of shBRCA1(knock down BRCA1). e Luciferase activity assay of mouse S100a9 promoter after 72 h transfection with PGL vector only, S100a9 promotor, S100a9 promotor with Brca1 cDNA, and S100a9 promotor with S100a9 cDNA in B477 and G600 cells. f S100A9 proteins in B477 mouse mammary epithelial cells and 231 cells with the expression of shBrca1 or shBRCA1, respectively by IF. g BRCA1 and S100A9 protein levels in B477 mouse WT mammary epithelial cells and 231 cells without or with the expression of shBRCA1(shBr/shBR). The data are expressed as means ± SD (a–e) and P values determined by unpaired two-tailed Student’s t test (a, c, d) and by one-way ANOVA followed by Tukey’s multiple comparisons (b) or two-way ANOVA (e). The experiments were independently repeated three times with similar results (a–g). Scale bar: white color, 20 μM. Source data are provided as a Source data file.

Article Snippet: Mouse Brca1 cDNA (Cat# MG212086) was purchased from Origene.

Techniques: Expressing, Control, Knockdown, Luciferase, Activity Assay, Transfection, Plasmid Preparation, Two Tailed Test

Journal: Nature communications

Article Title: S100A9-CXCL12 activation in BRCA1-mutant breast cancer promotes an immunosuppressive microenvironment associated with resistance to immunotherapy.

doi: 10.1038/s41467-022-29151-5

Figure Lengend Snippet: Fig. 4 Tumor permissive microenvironment in Brca1 MT mammary glands. a S100a9/S100a8 mRNA expression in the subpopulations of luminal and stromal cells of WT 4-month mammary gland (WTV4MG) and MT 4-months-old virgin mammary gland (MTV4MG) (n = 3 mice). b Protein level of S100a9 in both WT (B477) and MT (G600) mammary epithelial cell lines and tumor tissues by Western blots (n = 3 individual experiment-up and n = 3 mice-down). c Co-staining of S100a9 (red) and CK18 (green) with antibodies on WTV4MG, MTV4MG, WTV6MG, and MTV6MG tissues (n = 3 pairs in each group, Scale bar: 20 μM). d The S100a9 and Arg1 positive cell populations by FACS analysis from the blood and mammary tissues of both WT and MT mice at 4-month and 6-month, respectively (FACS gating strategies see in Supplementary Fig. 8c, n = 3 mice/ group). e Co-staining with S100a9 (red) and CD206 (green) antibodies (left panel) and co-staining with S100a9 (red) and CK18 (green) antibodies (right panel) on tumor-adjacent tissues by IF (40X confocal microscope, Scale bar: 20 μM.) (n = 3 mice and 3 individual experiment). f Secreted S100a9 proteins (left) from both tumor cell and MDSC cells in tumor-adjacent mammary gland (n = 3 mice) and present in the supernatant of cultured cancer cells (right) (n = 3 individual experiment, Scale bar: 10 μM). g Protein levels of S100a9, TGF-β, and Il-10 in mammary gland tissues of both WT and Brca1 MT mice at 4-month (n = 3 mice). h Protein levels of S100a9, TGF-β, and IL-10 in mammary tissues of both WT and Brca1 MT mice at 6-month (n = 3 mice). The data are expressed as means ± SD (a) and P values determined by unpaired two-tailed Student’s t test. The experiments were independently repeated three times with similar results (a, b). Source data are provided as a Source data file.

Article Snippet: Mouse Brca1 cDNA (Cat# MG212086) was purchased from Origene.

Techniques: Expressing, Western Blot, Staining, Microscopy, Cell Culture, Two Tailed Test

Journal: Nature communications

Article Title: S100A9-CXCL12 activation in BRCA1-mutant breast cancer promotes an immunosuppressive microenvironment associated with resistance to immunotherapy.

doi: 10.1038/s41467-022-29151-5

Figure Lengend Snippet: Fig. 8 Schematic for the mechanisms by which BRCA1 deficiency in breast cancer formation and improves responses to anti–PD1 antibody. In BRCA1 deficiency epithelial cells, S100A9 expression level constantly increases in early stages and secrets out to recruit and activate MDSCs, which creates an immunosuppression microenvironment by inhibit expansion and activation of T cells. This process can be further enhanced by CXCL12 that positively regulated by S100A9 and form a positive feedback loop. And this immunosuppression microenvironment is beneficial for tumor growth. The inhibitors of S100A9 and CXCL12, Tasquinimod and AMD3465, combine with anti-PD1 antibody can rescue the immunosuppression microenvironment and repress tumor growth. Green arrows indicate a decrease; red arrows indicate an increase.

Article Snippet: Mouse Brca1 cDNA (Cat# MG212086) was purchased from Origene.

Techniques: Expressing, Activation Assay