Fig. 5: Decitabine and C29 act in concert to suppress BC tumor growth in vivo.

a Schematic illustrating the establishment of a mouse BC CDX model from NIC-5231 cells for pharmacological drug testing in a pre-clinical setting. b Representative images of tumors extracted at endpoint (20 days post-treatment) from mouse mammary fat pads illustrating the effect of the different drug regimens on tumor size. c Tumor growth curves following administration of C29 (10 mg/kg), 5-azadC (1 mg/kg) or a combination of both drugs (n = 5 for each group). d Immunoblots of DNMT1 and ERRα in tumors after a 20-day drug regimen (n = 3 samples per group are shown). Tubulin is shown as a loading control. e Relative quantification of 5-methylcytosine levels in tumors after 20 days of treatment (n = 5 per group). f Computational pipeline developed for the identification of genes of interest from the RRBS analysis on tumors of mice treated with the different drug regimens. g Venn diagram representing the overlap of the number of DMRs in gene promoters after treatment with C29, 5-azadC or a combination of both drugs. A heatmap of the 51 transcription factors found with differential promoter methylation after C29 and 5-azadC co-treatment is shown. Red represents promoter hypermethylation, while blue designates promoter hypomethylation. The red arrow points at the transcription factor IRF4. h Subset of TF motif enrichment analysis performed on the total list of DMRs identified following treatment with C29, 5-azadC or a combination of both drugs. The IRF4 motif (boxed in red) was specifically enriched in promoters with DMRs of tumors co-treated with C29 + 5-azadC. Data shown in (c) and (e) represent means ± SEM. **p < 0.01, ***p < 0.001; Student’s t test.