Fig. 4: Assessment of the hypoxia/HIF-1α/adenosine/A2AR axis in tumour organoids.
From: Engineering micro oxygen factories to slow tumour progression via hyperoxic microenvironments
a Confocal imaging of hypoxic status. HepG2 and MCF-7 cells were incorporated in matrigel to prepare spheroids. After 7 days incubation, the constructed spheroids were exposed to 3600/mL PMCs and received 300 mW/cm2 NIR radiation for three intervals. The treated spheroids were subjected to staining by HypoxyprobeTM−1 Plus Kit (green) and Hoechst 33342 (blue). b HIF-1α and c adenosine productions in cancer cell spheroids (n = 3). The cells in spheroids were collected and lysed for HIF-1α or adenosine detection by ELISA and LC-MS, respectively. d Representative images and e quantification of IFN-γ expression in T cells by flow cytometry analysis (n = 3). T cells were treated with 50 ng/ml PMA, and 1 µL/mL Golgi inhibitor for 18 h with/without NIR-PMC for three intervals. The treated cells were fixed, permeabilized and stained by anti-IFN-γ-FITC for flow cytometry analysis. f Impacts of hyperoxia on the cytolytic activity of NK cells (n = 3). The sorted NK cells (2 × 105 cells/well) were mixed with Yac-1 cells at the ratios of 1:1 or 2:1 in 24-well plates, and co-cultured with/without PMCs, followed by three intervals of NIR exposure. After 6 h incubation, the cell mixtures were collected to lyse Yac-1 cells. The luminescence of cell lysates was measured by a Microplate reader. Data are expressed as mean ± SD. **p < 0.01 and ***p < 0.001 compared to Ctrl cells by two-tailed Student t-test. Source data are provided as a Source data file.
