Project description:ChIP sequencing for E2F1, E2F3A and E2F3B in mouse embryonic fibroblasts E2F1, E2F3A and E2F3B ChIPs were performed in mouse embryonic fibroblasts stably overexpressing the proteins. ChIP DNA was processed to generate libraries and then sequenced using the Illumina platform.
Project description:E2FÂs are regulators of the cell cycle and are involved in development. In this study we examine transcriptional changes occurring the liver in E2f1 (1KI) and E2f3b (3bKI) knock in mice. These mice have E2f1 or E2f3b knocked into the E2F3a locus resulting in loss of E2f3a and expression of E2f1 or E2f3b from the E2f3a locus as originally described In Tsai et. al., Nature 2008. Microarrays were used to evaluate transcriptional changes due to alterations in E2F expression during liver development. Affymetrix microarrays were performed using RNA samples from 4 weeks (1 month) old livers from wildtype (wt), E2F3a knockout (3aKO) E2f1 knock-in (1KI) and E2f3b knock-in (3bKI) mice.
Project description:E2Fs are regulators of the cell cycle and are involved in development. In this study we examine transcriptional changes occurring the liver in E2f1 (1KI) and E2f3b (3bKI) knock in mice. These mice have E2f1 or E2f3b knocked into the E2F3a locus resulting in loss of E2f3a and expression of E2f1 or E2f3b from the E2f3a locus as originally described In Tsai et. al., Nature 2008. Microarrays were used to evaluate transcriptional changes due to alterations in E2F expression during liver development.
Project description:E2Fâs are regulators of the cell cycle and are involved in development and hepatocellular carcinoma. In this study we examine transcriptional changes occurring the liver in E2f1 (1KI) and E2f3b (3bKI) knock in mice. These mice have E2f1 or E2f3b knocked into the E2F3a locus resulting in loss of E2f3a and expression of E2f1 or E2f3b from the E2f3a locus as originally described In Tsai et. al., Nature 2008. Microarrays were used to evaluate transcriptional changes in spontaneous hepatocellular carcinoma that arose due to alterations in E2F expression. Affymetrix microarrays were performed using mRNA samples from 12 month old livers from wildtype (wt), E2F3a knockout (3aKO) E2f1 knock-in (1KI) and E2f3b knock-in (3bKI) mice. Wt and 3aKO samples were from normal liver and 1KI and 3bKI samples were from spontaneously occurring hepatocellular carcinoma.
Project description:E2F’s are regulators of the cell cycle and are involved in development and hepatocellular carcinoma. In this study we examine transcriptional changes occurring the liver in E2f1 (1KI) and E2f3b (3bKI) knock in mice. These mice have E2f1 or E2f3b knocked into the E2F3a locus resulting in loss of E2f3a and expression of E2f1 or E2f3b from the E2f3a locus as originally described In Tsai et. al., Nature 2008. Microarrays were used to evaluate transcriptional changes in spontaneous hepatocellular carcinoma that arose due to alterations in E2F expression.
Project description:To investigate the impact of E2F1 loss on glutamine metabolism we isolated mouse embryonic fibroblasts from E2f1-wildtype (wt) and E2f1-knockout (ko) mice We then performed gene expression profiling analysis using data obtained from RNA-seq of 2 different treatment conditions (standard growth media vs media containing glutamine but depleted of glucose ).
Project description:Long-lasting changes in neuronal gene expression in brain reward regions, including the nucleus accumbens (NAc), contribute to the persistent functional changes in the addicted brain. Our group and others have demonstrated that altered expression or activity of several candidate transcription factors in NAc regulates drug responses. In a recent study from our group (Feng et al., 2014), involving large-scale genome-wide datasets, E2F3 was predicted as a prominent upstream regulator of cocaine-induced changes in gene expression and alternative splicing. Here, we show that E2F3a, but not E2F3b, expression in NAc regulates cocaine-induced locomotor and reward behavior. Furthermore, we demonstrate that E2F3a overexpression recapitulates a considerable portion of genome-wide transcriptional profiles and alternative splicing induced by cocaine administration. We further validate functional binding of E2F3a at several target genes following cocaine exposure. These novel findings support a crucial role for E2F3a in the regulation of cocaine-elicited behavioral states and molecular mechanisms.
Project description:We have found three specific combinations of two transcription factors, comprising Hnf4alpha plus Foxa1, Foxa2 or Foxa3, could convert mouse embryonic fibroblasts (MEFs) into cells that closely resemble hepatocytes in vitro. Then we used ChIP-seq to explore the targets of the transduced Foxa and Hnf4alpha proteins in MEFs.
Project description:To identify genes specifically activated by deregulated E2F, we examined gene expression profiles using human normal fibroblasts (HFFs), which were starved of serum and re-stimulated with serum, over-expressed with E2F1 or expressed with adenovirus E1a to forcedly inactivate RB. To identify genes activated by growth stimulation, human normal fibroblasts (HFFs) were starved of serum for 48 hr, infected with control virus, restimulated with serum or left serum-starved for 18 hr and harvested. To identify genes actived by deregulated E2F, HFFs were starved of serum for 48 hr, infected with control virus, adenovirus expressing E2F1 or adenovirus E1a, further cultured in the absence of serum for 18 hr and harvested. Gene expression profiles of ecach sample were examined by DNA microarray.