αccl2 antibody  (Bio X Cell)

Journal: Nature cancer

Article Title: EZH2 inhibition remodels the inflammatory senescence-associated secretory phenotype to potentiate pancreatic cancer immune surveillance

doi: 10.1038/s43018-023-00553-8

Figure Lengend Snippet: a, KEGG pathway analysis of RNA-seq data showing enriched pathways in shEzh2 compared to shRen KPC1 orthotopic PDAC tumor cells FACS sorted from C57BL/6 female mice treated with trametinib (1mg/kg) and palbociclib (100mg/kg) for 2 weeks (n=5-6 per group). b, Heatmap of RNA-seq analysis of SASP gene expression in tumor cells FACS sorted from KPC1 orthotopic PDAC tumors harboring indicated shRNAs and treated as in (a) (n=5-6 per group). c, qRT-PCR analysis of Ccl2 expression in KPC1 PDAC cells engineered to overexpress (O/E) a Ccl2 cDNA or Empty control vector (n=3 per group). A.U., arbitrary units. d, NK cell migration assay in the presence of conditioned media from KPC1 PDAC cells engineered to overexpress Ccl2 or Empty vector and treated with vehicle or trametinib (25nM) and palbociclib (500nM) for 8 days (n=3 per group). e, Flow cytometry analysis of NK cell numbers in KPC1 orthotopic PDAC tumors expressing control Empty or Ccl2 vectors following treatment as in (b) (e, Empty V, n=4; Empty TP, n=9; CCL2 O/E, n=11 and CCL2 O/E/TP, n=12 independent mice). Data represents pool of 3 independent experiments. f, Kaplan-Meier survival curve of mice with KPC1 orthotopic PDAC tumors expressing control Empty (left) or Ccl2 (right) vectors treated with vehicle, combined trametinib (1mg/kg) and palbociclib (100mg/kg), and/or an NK1.1 depleting antibody (PK136; 250 μg) (f, Empty V, n=12; Empty TP, n=14; Empty TP+αNK1.1, n=13; CCL2O/E V, n=10; CCL2 O/E TP, n=11 and CCL2 O/E TP+αNK1.1, n=12 independent mice). Data represents pool of 2 independent experiments. g, Flow cytometry analysis of NK cell numbers in shEzh2 KPC1 orthotopic PDAC tumors following treatment with vehicle, combined trametinib (1mg/kg) and palbociclib (100mg/kg), and/or a CCL2 depleting antibody (2H5; 200 μg) for 2 weeks (g, V, n=3; αCCL2, n=3; TP, n=7 and TP αCCL2, n=9 independent mice). Data represents pool of 2 independent experiments. h, Waterfall plot of the response of shEzh2 KPC1 orthotopic PDAC tumors to treatment as in (g) (h, shEzh2 TP, n=22 and shEzh2 TP αCCL2, n=23 independent mice). Data represents pool of 3 independent experiments. i, Kaplan-Meier survival curve of mice with shEzh2 KPC1 orthotopic PDAC tumors treated as in (g) (i, shEzh2 V, n=13; shEzh2 TP, n=15 and shEzh2 TP αCCL2, n=10 independent mice) Values for shEzh2 V and T/P treated cohorts are the same displayed in Figs. 5h and ​and5i.5i. Dotted line indicates when mice were taken off of treatment. Data represents pool of 2 independent experiments. j, Flow cytometry analysis of CD4+ and CD8+ T cell numbers in shEzh2 KPC1 orthotopic PDAC tumors following treatment with vehicle, combined trametinib (1mg/kg) and palbociclib (100mg/kg), and/or a CXCR3 depleting antibody (CXCR3-173; 200 μg) for 2 weeks (j, V, n=8; αCXCR3, n=5; TP, n=8; and TP+αCXCR3, n=12 independent mice). Data represents pool of 2 independent experiments. k, Waterfall plot of the response of shEzh2 KPC1 orthotopic PDAC tumors to treatment as in (j) (k, V, n=11; αCXCR3, n=7; TP, n=13; and TP+αCXCR3, n=15 independent mice). Data represents pool of 2 independent experiments. P values in a were calculated using two-sided, hypergeometric test, c,d,e,g,h,j,k using two-tailed, unpaired Student’s t-test, and f and i using log-rank test. Error bars, mean ± SEM.

Article Snippet: For neutralization of chemokine signaling, mice were injected IP with an αCCL2 (200 μg; 2H5, BioXcell) or αCXCR3 (200 μg; CXCR3-173, BioXcell) antibody twice per week.

Techniques: RNA Sequencing, Gene Expression, Quantitative RT-PCR, Expressing, Control, Plasmid Preparation, Cell Migration Assay, Flow Cytometry, Two Tailed Test

Journal: Nature cancer

Article Title: EZH2 inhibition remodels the inflammatory senescence-associated secretory phenotype to potentiate pancreatic cancer immune surveillance

doi: 10.1038/s43018-023-00553-8

Figure Lengend Snippet: a, IHC staining of KPC1 orthotopic PDAC tumors harboring shRen or shEzh2 shRNAs treated with vehicle or combined trametinib (1mg/kg) and palbociclib (100 mg/kg) (T/P) for 2 weeks. Quantification of blood vessels per field are shown on inset (a, shRen V, n=4; shRen TP, n=4; shEzh2 V, n=2; shEzh2 TP, n=4 independent tumors). Scale bar, 50μm. b-c, Flow cytometry analysis of NK cell activation markers (b) and CD4+ and CD8+ T cell numbers (c) in KPC1 orthotopic PDAC tumors expressing control Empty or Ccl2 vectors and treated as in (a) (b-c, Empty V, n=4; Empty TP, n=9; Empty CCL2O/E V, n=11; CCL2 O/E TP, n=12 independent mice). Data represents pool of 3 independent experiments d, Flow cytometry analysis of CD4+ and CD8+ T cell numbers in shEzh2 KPC1 orthotopic PDAC tumors following treatment with vehicle, combined trametinib (1mg/kg) and palbociclib (100mg/kg), and/or a CCL2 depleting antibody (2H5; 200 μg) for 2 weeks (d, shEzh2 V, n=3; shEzh2 αCCL2, n=3; shEzh2 TP, n=7; shEzh2 TP+αCCL2, n=9 independent mice). Data represents pool of 2 independent experiments. e, Flow cytometry analysis of NK cell numbers in shEzh2 KPC1 orthotopic PDAC tumors following treatment with vehicle, combined trametinib (1mg/kg) and palbociclib (100mg/kg), and/or a CXCR3 depleting antibody (CXCR3-173; 200 μg) for 2 weeks (d, V, n=8; TP, n=8; αCCXR3, n=5; TP+αCCXR3, n=12 independent mice). Data represents pool of 2 independent experiments. P values in a-e were calculated using two-tailed, unpaired Student’s t-test. Error bars, mean ± SEM.

Article Snippet: For neutralization of chemokine signaling, mice were injected IP with an αCCL2 (200 μg; 2H5, BioXcell) or αCXCR3 (200 μg; CXCR3-173, BioXcell) antibody twice per week.

Techniques: Immunohistochemistry, Flow Cytometry, Activation Assay, Expressing, Control, Two Tailed Test