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RNA-seq data for lineage negative, Sca1 positive, and cKit positive hematopoietic stem/progenitor cells from H3.3K36M, H3.3K9M, or rtTA control mice

ABSTRACT: We analyzed gene expression patterns in LKS cells using RNA-seq after one week of mutant histone induction. We chose this time point to minimize secondary effects and because methylation at both H3K9 and H3K36 is maximally depleted by that time. Compared to rtTA control, 150 and 364 genes were differentially expressed in H3K9M and H3K36M expressing LKS cells, respectively. Principal component analysis indicated that biological replicates were highly consistent and expression profiles for H3K9M, H3K36M, and rtTA control were distinct. In general, gene expression changes following H3K9M induction were subtle, while induction of H3K36M led to more robust differences in expression patterns.

ORGANISM(S): Mus musculus

PROVIDER: GSE103521 | GEO | 2020/09/04

REPOSITORIES: GEO

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Project description:Increasing evidence suggests that microRNAs may play important roles in regulating self-renewal and differentiation in mammalian stem cells (SCs). Here, we explore this issue in skin. We first characterize microRNA expression profiles of skin SCs versus their committed proliferative progenies and identify a microRNA subset associating with “stemness”. Of these, miR-125b is dramatically downregulated in early SC-progeny. We engineer an inducible mice system and show that when miR-125b is sustained in SC-progenies, tissue balance is reversibly skewed towards stemness at the expense of epidermal, oil-gland and HF differentiation. Using gain-and-loss of function in vitro, we further implicate miR-125b as a repressor of SC differentiation. In vivo, transcripts repressed upon miR-125b induction are enriched >700% for predicted miR-125b targets normally downregulated upon SC-lineage commitment. We verify some of these miR-125b targets, and show that Blimp1 and VDR in particular can account for many tissue imbalances we see when miR-125b is deregulated. We used microarrays to compare the global miRNA expression profile of P4 stage hair follicle ORS cells from DTG (K14-rtTA,TRE-miR-125b) and control littermates. Hair follicle cells were isolated from P4 Backskin (Dox since P3 for 24hrs) of DTG (K14-rtTA/TRE-miR-125b/K14-H2BGFP), TRE (TRE-miR-125b/K14-H2BGFP), KrtA (K14-rtTA/K14-H2BGFP) as following: interfollicular epidermis sheet was pealed from hair follicle & dermis after dispase treatment.The hair follicle & dermis were first digested by collagenase (Sigma). Intact hair follicles were separated from dermal cells by low speed spinning (20g). The hair follicles were then digested by Trypsin and filtered by 40 µm cell strainers. The isolated hair follicle cells were FACS sorted. During FACS, cells were first gated against CD34 (endothelial cells), CD45 (immune cells), CD114 (melanocytes) and DAPI (dead cells). ORS cells were sorted from the remaining cells as α6HiGFPHi.

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RNA-seq data for lineage negative, Sca1 positive, and cKit positive hematopoietic stem/progenitor cells from H3.3K36M, H3.3K9M, or rtTA control mice

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