Dataset Information

HEK293T 15xGly/99xGly immunoprecipitation-mass spectrometry

ABSTRACT: In order to determine the protein composition of polyglycine aggregates, clonal HEK293T cell lines harboring doxycycline (DOX)-suppressible 15xGly-eGFP-3xHA or 99xGly-eGFP-3xHA constructs were grown either in standard media for 14 days (induction of transgene) or maintained in DOX (suppression of transgene).

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell, Cell Culture

DISEASE(S): Disease Free

SUBMITTER: Raghu Chivukula

LAB HEAD: Raghu R. Chivukula

PROVIDER: PXD046400 | Pride | 2025-07-18

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

Items per page:

1 - 5 of 37

Similar Datasets

Project description:We aim to understand the role that Cdx2 plays in specifying the rostro-caudal identity of differentiating motor neurons. We find that expressing Cdx2 in combination with FGF signaling is sufficient to produce motor neurons with a more caudal identity. ChIP-seq analysis of Cdx2 finds that it binds extensively throughout the Hox regions in progenitor motor neurons. Analysis of polycomb-associated chromatin over Hox regions in the subsequently generated motor neurons finds that Cdx2 binding corresponds to chromatin domains encompassing de-repressed caudal Hox genes. These results suggest a direct role for Cdx2 in specifying caudal motor neuron identity. ChIP-seq studies: We characterize the binding of Cdx2 in progenitor motor neurons using a V5 tagged doxycycline inducible Cdx2 ESC line (iCdx2). Progenitor motor neurons were generated after 4 days of in vitro differentiation of mouse embryonic stem cells using retinoic acid (RA) and hedgehog (Hh) signaling exposure at day 2. On day 3, the cells are exposed to Dox with and without accompanying FGF signaling. The genome-wide binding of the induced Cdx2 transcription factor is profiled using ChIP-seq with an anti-V5 antibody. An appropriate whole-cell extract control experiment for these ChIP-seq experiments is also included. We also examine the effect of induced Cdx2 expression on polycomb-associated chromatin structure in the resulting cellular populations by profiling the H3K27me3 chromatin mark using ChIP-seq. H3K27me3 experiments were performed after 5 days of in vitro differentiation using cells exposed to either: 1) RA & Hh to derive progenitor motor neurons, followed by Dox & FGF; 2) Dox & FGF alone; or 3) RA and Hh alone. There are 6 Illumina sequencing datasets included in this submission: two biological replicates of iCdx2 ChIP-seq in the presence of FGF; one sample of iCdx2 ChIP-seq in the absence of FGF; one H3K27me3 ChIP-seq in the presence of RA, Hh, Dox, and FGF; one H3K27me3 ChIP-seq in the presence of Dox and FGF; and one H3K27me3 ChIP-seq in the presence of RA and Hh.

			Action	DRS
	220318_15_3raghu-1A.raw	Raw
	220318_16_3raghu-1B.raw	Raw
	220318_17_3raghu-1C.raw	Raw
	220318_18_3raghu-1D.raw	Raw
	220318_19_3raghu-1E.raw	Raw

Dataset Information

HEK293T 15xGly/99xGly immunoprecipitation-mass spectrometry

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets