Fig. 5: CAF-CD73 expression is dynamically upregulated via the ADO-A_2B pathway.

a The relative abundancy and distribution of CAFs in EG7 tumors of various sizes were evaluated via ER-TR7 (green) staining using a computer-assisted software, MetaMorph. Data depict mean ± SEM. b, c The CAF abundancy and CAF-CD73 levels in EG7 of various sizes were analyzed via FACS. d EG7-CAFs were treated with 100 μM ADO for 6 and 24 h and their Nt5e (Cd73) expression was determined via quantitative real-time RT-PCR. e EG7-CAFs were treated with 100 μM ADO in the absence or presence of A_2A or A_2B antagonists, ZM241385 or PSB1115, respectively. Relative CD73 expression on CAFs was determined via FACS. f EG7 tumor-bearing WT mice were treated with daily i.p. injection of 1 mg kg⁻¹ body weight of PSB1115 or vehicle (DMSO) as a control. CD73 expression on CAFs were examined via FACS. g WT and A_2b^null mice were inoculated with EG7 tumors and their CAF-CD73 expression from tumors of similar sizes was compared. h EG7 tumor-bearing WT mice were treated with daily i.p. injection of 1 mg kg⁻¹ body weight of PSB1115 or vehicle starting from 5-day post-tumor inoculation followed with i.t. injections of 100 μl of 20 μM nutlin at days 6 and 8. i EG7 tumor-bearing A_2b^null mice were treated with i.t. injections of 100 μl of 20 μM nutlin at days 6 and 8 post-tumor establishment. CD73 expression on CAFs from WT h and A_2b^null i mice were analyzed via FACS and compared to those without nultin-treatment (n = 5). Error bars depict mean ± SEM. p values were determined via two-tail unpaired Student’s t-test a, d–i. Linear regression analysis was performed to determine the correlation between the tumor size and CAF% or CD73⁺CAF% in b, c, respectively. All experiments were repeated at least two times. Source data are provided in the Source Data file.