Additional file 2: Figure S1. of Nrf2 status affects tumor growth, HDAC3 gene promoter associations, and the response to sulforaphane in the colon

Validation of gene array data in mouse colon tumors and normal colon. a–d Relative mRNA expression of selected genes normalized to the β-actin gene (Actb) in normal colonic mucosa, and e–h in DMH-induced colon tumors. Data = mean ± SD (n = 3). Red arrow, p16. Figure S2. HDAC3 interactions with Esr1 are unaffected by SFN. Primers were designed to interrogate the promoter region adjacent to the transcriptional start site of Esr1, a gene strongly overexpressed in DMH-induced colon tumors (see Fig. 5b). ChIP data = mean ± SD from three independent ChIP assays. Esr1 served as a negative control for the ChIP assays with p16 (see Fig. 6f). Figure S3. SFN altered HDAC3 and p16 expression in mouse splenocytes. Mice (n = 3) were administered a single oral gavage of 200 μmol SFN, and mononuclear cells were isolated from mouse spleen at 6 h, as reported earlier [12]. a Whole cell lysates were immunoblotted for HDAC3, p16, and AcH4K12 with densitometry data normalized to β-actin. b A colon tumor from DMH-treated mice was included as a reference control. Figure S4. No change in the expression of genes involved in DMH activation (Cyp2E1) and DNA repair (Mgmt). Relative mRNA expression levels were normalized to the β-actin gene, Actb. Data = mean ± SD (n = 3). Figure S5. Working model for Nrf2 status impacting tumor growth, HDAC3 levels, and p16 induction. (PPTX 276 kb