Journal: Science Advances

Article Title: Immunosuppressive macrophages determine the effect of cellular senescence on tumor progression

doi: 10.1126/sciadv.adx2988

Figure Lengend Snippet: ( A ) Tumor samples from mice treated with vehicle or AP21967 at 9 weeks were homogenized, and the levels of the indicated cytokines were analyzed using a cytokine array. The log 2 FC (AP21967/vehicle) for each cytokine is presented. ENA-78, epithelial-derived neutrophil-activating peptide 78; GROα, growth-regulated oncogene-alpha. ( B ) Levels of CCL2 and CXCL1 were quantified by ELISA from the same tumor homogenates as in (A). ( C ) UMAP plots showing the expression of the most up-regulated cytokines (CCL2, CCL3, and CCL7) and their receptors (CCR1, CCR2, and CCR5) in different cell populations. Relevant cell clusters are indicated. N, neutrophils; MØ, macrophages; E, endothelial cells; F, fibroblasts. ( D ) Flow cytometry analysis of myeloid cells purified from tumors obtained from SuSe mice treated with vehicle or AP21967. We identified TAMs and cells positive for the myeloid marker CD11B and for the macrophage marker F4/80. ( E ) The same cells as in (D) were cultured for 24 hours—in the presence of vehicle or anti-CCL2—and the presence of CCL2 in the conditioned medium was determined by ELISA. ( F ) Schematic representation of the specificities of receptors for CCL2, CCL3, and CCL7. ( G ) Flow cytometry analysis of CCR2 in the same cells as in (D). FSH-H, forward scatter height.

Article Snippet: For selective CCL2 blocking, 4-week-old PyMT/SuSe mice were treated every 3 days with anti-CCL2 antibody (#BE0185, BioXCell,) at a concentration of 2 μg/g of body weight.

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Expressing, Flow Cytometry, Purification, Marker, Cell Culture

Journal: The Journal of Clinical Investigation

Article Title: Comprehensive genomic profiling of triple-negative breast cancer metastases identifies role of PKD1 in immunotherapy resistance

doi: 10.1172/JCI188989

Figure Lengend Snippet: ( A ) Schematic of the treatment with anti–PD-1 and/or anti-CCL2 in mice from the PKD1-NC and PKD1-OE groups. ( B ) Tumor growth curves of mice in the PKD1-NC and PKD1-OE groups treated with anti–PD-1 and/or anti-CCL2 antibodies. P values were calculated via 2-tailed unpaired Student’s t tests. ( C ) Multiplex immunofluorescence results of CD45 + CD8 + cells. Left: Representative multiplex immunofluorescence images. Right: Quantitative analysis across treatment groups. Statistical comparisons were performed using Dunnett’s test, with the vehicle group designated as the reference control. ( D ) Multiplex immunofluorescence results of F4/80 + CD86 + cells. Left: Representative multiplex immunofluorescence images. Right: Quantitative analysis across treatment groups. P values were assessed using Dunnett’s test. ( E ) Heatmap showing immune cell infiltration in the tumor immune microenvironment across treatment groups (vehicle, anti–PD-1, anti-CCL2, and combined), based on bulk RNA-seq of murine TNBC. P values were calculated using the Kruskal-Wallis test. CCL2, C-C motif chemokine ligand 2; TNBC, triple-negative breast cancer; NC, negative control; OE, overexpression. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Mice assigned to immunotherapy were given 5 intraperitoneal injections of 200 μg InVivoMAb anti-mouse PD-1 (Bio X Cell) or/and anti-mouse CCL2-InVivo (Selleck) every third day.

Techniques: Multiplex Assay, Immunofluorescence, Control, RNA Sequencing, Negative Control, Over Expression