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Effect of HES1 overexpression on gene expression of adipose stem cells [RNA-seq]

ABSTRACT: Effect of HES1 overexpression on gene expression of adipose stem cells [RNA-seq]

PROVIDER: PRJNA1124895 | ENA |

REPOSITORIES: ENA

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Effect of HES1 overexpression on gene expression of adipose stem cells [RNA-seq]

Project description:In the current investigation, an RNA-seq analysis was conducted to examine the differentially expressed genes in old ADSCs (O-ADSCs) following the overexpression of HES1. The findings indicated that 448 genes exhibited increased levels, while 865 genes showed decreased levels in the oe-HES1 O-ADSCs compared to the oe-NC O-ADSCs. The GO analysis revealed that these genes are primarily localized in the nucleus and cytoplasm, and are involved in activities such as protein binding and DNA binding. Furthermore, they are associated with processes related to cell adhesion and the regulation of transcription by RNA polymerase II. The KEGG analysis indicated enrichment in pathways such as the NOD-like receptor signaling pathway, TNF signaling pathway, IL-17 signaling pathway, and PI3K-Akt signaling pathway.

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Project description:A human colon carcinoma-derived cell line LS174T was modified to overexpress Hes1, a bHLH-type transcription factor, upon doxycycline addition (designated as LS174T-tetON-Hes1 cells), using the T-rex system (Invitrogen). We have previously shown that these cells can overexpress Hes1 under the control of CMV promoter (Zheng et al, Inflamm Bowel Dis, 17;2251-2260, 2011), and the amont of the overexpressed Hes1 protein reaches to the maximal level in early as 3 hours from doxycycline addition (100ng/ml), which persists for up to 24 hours. In the present experiment, LS174T-tetON-Hes1 cells were treated with recombinant human IL-22 (20ng/ml) alone, doxycycline alone (100ng/ml), co-treated by both IL-22 and doxycycline, or left untreated (Control) for 24 hours, and subjected for analysis. Experiment was done using a modified sub-line of LS174T cell (LS174T-tetON-Hes1 cells), in which overexpression of Hes1 can be induced by a Doxycycline dependent manner. Four-condition experiment, Control vs. IL-22 stimulation, Control vs Doxycycline addition (Hes1 overexpression) and Control vs IL22 stimulation+Doxycycline addition (Hes1 overexpression). Biological replicates: One for each condition, independently grown and harvested. One replicate per array.

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