Figure 2 from Ammonia Suppresses the Antitumor Activity of Natural Killer Cells and T Cells by Decreasing Mature Perforin

<p>Ammonia concentration is increased in cancer cell–conditioned medium and TIF. Ammonia concentration in the tumor-conditioned medium, collected after 48 hours of incubation, was measured using a Dimension Ammonia assay (Siemens; <i>n</i> = 3). Cells were cultured at different densities: lymphoma (0.5 × 10⁶, 1.0 × 10⁶, 1.5 × 10⁶/mL; <b>A</b>), multiple myeloma (1 × 10⁶, 2 × 10⁶, 4 × 10⁶/mL; <b>B</b>), and breast cancer cell lines (0.5 × 10⁶, 1 × 10⁶, 1.5 × 10⁶/mL; <b>C</b>). In <b>A–C</b>, empty medium incubated for 48 hours (without the cells) at 4°C or 37°C is presented as a control. <b>D,</b> Schematic presentation of the TIF and SCF isolation from mice. TIF was collected from tumors not exceeding 1,500 mm³. SCF was isolated at the same time from the contralateral tight as a control tissue fluid. <b>E–G,</b> The concentration of ammonia in TIF and SCF isolated from Raji tumor–bearing NSG (<i>n</i> = 5; <b>E</b>), MM.1s tumor–bearing SCID mice (<i>n</i> = 5; <b>F</b>), and breast cancer–bearing mice (EMT6 in BALB/c mice, <i>n</i> = 5; E0771 in BALB/c mice, <i>n</i> = 5; 4T1 in BALB/c mice; <i>n</i> = 5; MCF7 in NSG mice, <i>n</i> = 4; MDA-MB-231 in NSG mice, <i>n</i> = 3; <b>G</b>). <i>P</i> values were calculated using a paired <i>t</i> test. Data show individual values and means ± SEM. <i>n</i> values are the number of biological replicates in <i>in vitro</i> experiments or the number of mice used to obtain the data. <b>D,</b> Created with BioRender.com. Winiarska, M. (2025) <a href="https://BioRender.com/x56a982" target="_blank">https://BioRender.com/x56a982</a>.</p>