Fig. 7: Adenosinergic antagonism is ineffective in modulating CAF-CD73 expression in the CAF-poor MC38 tumors.

a MC38 tumor-bearing mice were treated with daily i.p. injection of A_2A antagonist ZM241385 or A_2B antagonist PSB1115 or anti-CD73 every other day. Tumor progression was examined every day. b, c The percentage of CD73^hi/+ CAFs (b) and MC38 tumors (c) in the TME following different treatments were analyzed via FACS. d MC38 tumor progression in WT mice receiving combination treatment of ZM243185 and PSB1115 in the absence or presence of anti-CD73 was examined every day. e, f The percentage of CD73^hi/+ CAFs (e) and MC38 tumors (f) in the TME following different treatments were analyzed via FACS. g Representative images of IHC staining of CD73 (red), α-SMA (green), and vimentin (white) revealed their levels and distribution within the MC38 TME. The percentage of a-SMA⁺ signal covering the entire TME was determined via the Inform software and presented to the left. h MC38 tumor established in WT mice were collected at different sizes for FACS analysis. The relative CAF abundancy and CD73^hi/+ CAFs within the MC38 TME in association with tumor size were evaluated via linear regression. i MC38 tumors were cultured with 100 μM ADO in the absence of presence of 10 mM ZM241385 or PSB115 for 48 h. Their Cd73 mRNA levels relative to that of b-actin was analyzed via real-time RT-PCR. j Representative images of IHC staining of CD73 (red), α-SMA (green), apoptotic cells (cleaved caspase-3⁺, cyan) and vimentin (white) levels and their distribution in the MC38 TME treated with or without ZM241385 and PSB115. a, d p values were determined via multiple t-tests where the tumor sizes at each time point were compared. b, c, e, f, g, i Error bars depict mean ± SEM. p values were determined via two-tailed unpaired Student’s t-test. g, h Scale bars, 50 μm. All experiments were repeated at least two times. Source data are provided in the Source Data file.