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: JOR Spine

Article Title: Explorative Study of Modulatory Effects of Notochordal Cell‐Derived Extracellular Vesicles on the IL ‐1β‐Induced Catabolic Cascade in Nucleus Pulposus Cell Pellets and Explants

doi: 10.1002/jsp2.70043

Figure Lengend Snippet: The effect of porcine notochordal cells (NC)‐derived EVs on pro‐catabolic factors in dog 3D NPC pellets. (A) IL‐6, (B) CXCL8/IL‐8, (C) CCL2, and (D) COX‐2 expression of dog pellets cultured in control medium, medium containing NC‐EVs, and NC‐EV‐depleted media, with and without canine IL‐1β, for 48 h. (E) IL‐6 release in the medium (dl: 15.6 pg), (F) prostaglandin E2 (PGE2) release in the medium (dl: 3.9 pg), and (G) immunohistochemistry for IL‐1β and IL‐1 receptor I (IL‐1RI) of dog pellets cultured for 7 days with corresponding immunopositive surface area (I and J, respectively). Pellets containing a pool of n = 7 dog nucleus pulposus donors were tested with individual porcine NCCM donors ( n = 5, pig 1–5). * p < 0.05, ** p < 0.01. Data are displayed as the grand mean with the individual donors plotted. Detection limit (dl) of the factors is indicated between brackets. The background levels of the EV‐ and EV‐depleted media alone (without pellet culture) were below the detection limit for all measured factors. ND: not detectable. Scale bar indicates 100 μm.

Article Snippet: Detection was performed with biotinylated anti‐swine CCL2 polyclonal antibody (0.1 μg/mL, 60 min, Kingfisher Biotech, PBB0089S‐050), high‐sensitivity streptavidin‐horseradish peroxidase (20 ng/mL, 30 min, Thermo Scientific, 21 134), and 3,3′,5,5′‐tetramethylbenzidine (TMB) substrate (30 min, Thermo Scientific, N301).

Techniques: Derivative Assay, Expressing, Cell Culture, Control, Immunohistochemistry