Fig. 4
Guadecitabine enhances IFNγ-mediated MHC-I expression and Cxcr3 ligands. a MMTV-neu tumors were resected 7 days after a 3-day in vivo treatment course with guadecitabine or diluent control. Tumor RNA was extracted and utilized for NanoString gene expression analysis using the PanCancer Immune Pathways codeset (>700 immune-related genes). Genes significantly altered (nominal p-value < 0.05) between the treatment groups are shown with a row-standardized z-scores in heatmap form. Association of altered genes with key pathways are color coded on the right. b MMTV-neu cells were cultured for 7 days with the indicated doses of guadecitabine, followed by 24 h treatment with IFNγ and evaluation by western blot analysis. c MMTV-neu cells were treated as in b but co-treated with or without BMS-345541 (during last 24 h, in tandem with IFNγ) and assessed by flow cytometry for MHC-I. Data represent the mean of four experiment replicates ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001; ns nonsignificant. d qRT-PCR for Cxcr3 ligands in MMTV-neu cells after 7 days of guadecitabine treatment at the indicated doses and 24 h treatment with IFNγ. Data represent the mean of 3 experiment replicates ± SEM. e Relative mRNA expression of Cxcr3 ligands in MMTV-neu cells after 7 days of treatment with guadecitabine (0.5 µg/mL) or diluent control, and 24 h exposure to 10 µM BMS-345541. Data represent the mean of 3 experiment replicates ± SEM
