Ctla 4

For sporadic CRC model (Cdx2-Cre⁺/Apc^F/WT mice), IL-17A, CTLA-4, and PD-1 neutralizing antibodies or isotype control antibodies (Bio X Cell) were i.p. injected at a dose of 100 μg per mouse every 3 days until sacrifice.
For the tamoxifen inducible model of tumorigenesis, antibodies (100 μg per mouse, every 3 days) were injected 1 day after the dose of tamoxifen until sacrifice.

Tumor-free 6- to 8-week-old female C57/BL6 mice were injected systemically with 300 pmol of AS141-SMG1 AsiC on days 1, 2, 3, 10, 14, and 15 following the same schedule as for antitumor experiments. 100 μg of CTLA-4 and 100 μg of PD-1 or 200 μg of 4-1BB (Clone 3H3; Bio X Cell) antibodies were intraperitoneally administered at days 1, 3, and 7. On day 20, mice were sacrificed, and spleen and liver were extracted. Spleen samples were weighed to study splenomegaly. Liver samples were processed for flow cytometry (see flow cytometry section of this materials and methods) and additionally used for paraffin embedding. 5-μm slices were stained with hematoxylin and eosin staining to detect tissue damage and immune cell infiltration.