Project description:We aim to determine if mice in our mouse colony had similar of different microbiomes. To do this, we perfromed 16S sequencing of stool from unifected mice of the gentotypes listed below. We also looked at how infection causes dysbiosis of the mircobiome, measuring 16S sequencing over a C.rodentium infection timecourse.
Project description:IL22 induces antimicrobial peptides which influnce microbiota. We used 16s rRNA gene sequencing (16s DNA-seq) to analyze the microbiota with Fc or IL-22Fc treatment.
Project description:Epigenetic gene silencing induced by aberrant DNA methylation is an important mechanism leading to the loss of key cellular pathways in tumorigenesis. Although DNA demethylating agents reverse DNA methylation and have significant cytostatic and cytotoxic effects, it remains elusive whether their effects on cellular phenotypes are solely on reactivation of hypermethylated genes. To address this issue, we constructed LNCaP-derived human prostate cancer cell lines that can pharmacologically induce the expression of fusion gene comprising NFkB transcriptional activation domain (AD) and methyl-CpG binding domain (MBD). Tetracycline induction of NFkB (AD)-MBD protein led to so-called methyl-CpG targeted transcriptional activation (MeTA), as demonstrated by the reactivation of hypermethylated genes such as MT1M, NEFH, and NEFM. The cell proliferation assay indicated that MeTA suppressed the growth of LNCaP cells. Furthermore, both flow cytometry and TUNEL assay clearly demonstrated that MeTA induced apoptosis. In order to search genes responsible for apoptosis, we performed gene expression microarray analysis of MeTA-uninduced and -induced LNCaP cells: Several tumor necrosis factor receptor superfamily (TNFRSF) genes upregulated in accordance with the induction of MeTA. These results suggest that DNA methylation confers cancer cells the ability to avoid apoptosis and MeTA may provide an efficient mean to analyze the change in cancer cell phenotypes by DNA methylation alterations. This is the first report showing that the reactivation of hypermethylated genes by the method other than DNMT inhibition induces growth arrest and apoptosis in cancer cells.
