Figure S5: Immunohistochemical staining of CD8+ T cell infiltration level in colon carcinoma tissues from 10 patients. The brown color indicates specific CD8+ T cells.

Supplemental Figure Legends

Supplementary Figure 1 from Nitric Oxide Inactivates the Retinoblastoma Pathway in Chronic Inflammation

Supplementary Fig. S1 from A role for macroautophagy in protection against 4-hydroxytamoxifen–induced cell death and the development of antiestrogen resistance

Supplementary Fig. S3 from A role for macroautophagy in protection against 4-hydroxytamoxifen–induced cell death and the development of antiestrogen resistance

Supplementary Figure 2 from Nitric Oxide Inactivates the Retinoblastoma Pathway in Chronic Inflammation

S2. cGMP-elevating agents regulate homeostasis in intestinal epithelium

Phosphodiesterase-5 inhibitors (PDE5i) are under investigation for repurposing for colon cancer prevention. A drawback to conventional PDE5i are their side-effects and drug–drug interactions. We designed an analog of the prototypical PDE5i sildenafil by replacing the methyl group on the piperazine ring with malonic acid to reduce lipophilicity, and measured its entry into the circulation and effects on colon epithelium. This modification did not affect pharmacology as malonyl-sildenafil had a similar IC50 to sildenafil but exhibited an almost 20-fold reduced EC50 for increasing cellular cGMP. Using an LC-MS/MS approach, malonyl-sildenafil was negligible in mouse plasma after oral administration but was detected at high levels in the feces. No bioactive metabolites of malonyl-sildenafil were detected in the circulation by measuring interactions with isosorbide mononitrate. The treatment of mice with malonyl-sildenafil in the drinking water resulted in a suppression of proliferation in the colon epithelium that is consistent with results previously published for mice treated with PDE5i. A carboxylic-acid-containing analog of sildenafil prohibits the systemic delivery of the compound but maintains sufficient penetration into the colon epithelium to suppress proliferation. This highlights a novel approach to generating a first-in-class drug for colon cancer chemoprevention.

Table S2. Scores of CD4+ and CD8+ peritumoral lymphocytes, and tumor GPR109A protein levels in 10 human CRC specimens.

Figure S1. Silencing STAT1 diminishes IFN-gamma-induced GPR109A expression in human colon carcinoma cells.

Supplementary Figures 1-5 from GPR109A Is a G-protein–Coupled Receptor for the Bacterial Fermentation Product Butyrate and Functions as a Tumor Suppressor in Colon

Supplementary Fig. S2 from A role for macroautophagy in protection against 4-hydroxytamoxifen–induced cell death and the development of antiestrogen resistance

Supplementary Figure 3 from Nitric Oxide Inactivates the Retinoblastoma Pathway in Chronic Inflammation

<p>Table S3. Differential expression of inflammatory genes in human colon carcinoma</p>

Supplementary Figure Legends 1-3 from Nitric Oxide Inactivates the Retinoblastoma Pathway in Chronic Inflammation

There are two supplementary figures. Figure S1 shows Western blot analysis of polyps from control and sildenafil treated mice, for cGMP signaling pathways (VASP, beta-catenin, AKT). Figure S2 shows RT-qPCR analysis of the mucosa and polyps from control and sildenafil treated animals for the expression of phosphodiesterases.

S1. Non-polyp tissues from Apc Min/+ mice have increased levels of proliferation.