Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Expression profiling of mouse CD4+ T cells following Klf1 retrovirus vector transfection

ABSTRACT: Krüppel-like factor 1 (Klf1) is a transcription factor that promotes β-globin synthesis in erythroid progenitors, and its role in immunological homeostasis is unknown. The full-length mRNA of mouse Klf1(NM-010635) was cloned into the pMIG retroviral vector, which contains an IRES-GFP cassette. Splenocytes from C57BL/6 mice were cultured for 48 h in the presence of Concanavalin A (ConA) (10 μg/mL) and IL-2 (50 μg/mL), and gene transduction was conducted. CD4+ T cells that were transfected with retroviral vectors were divided into 3 fractions according to GFP fluorescence (strongly positive, weakly positive and negative) using a FACSVantage flow cytometer. Total RNA was then extracted. Sequence libraries were prepared according to standard methods using Smart-seq2 (Illumina), and they were analyzed with a next generation sequencer, a MiSeq system (Illumina).

INSTRUMENT(S): Illumina MiSeq

ORGANISM(S): Mus musculus

SUBMITTER: Shuzo Teruya

PROVIDER: E-MTAB-6778 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Egr2-independent, Klf1-mediated induction of PD-L1 in CD4+ T cells.

Teruya Shuzo S Okamura Tomohisa T Komai Toshihiko T Inoue Mariko M Iwasaki Yukiko Y Sumitomo Shuji S Shoda Hirofumi H Yamamoto Kazuhiko K Fujio Keishi K

Scientific reports 20180504 1

Programmed death ligand 1 (PD-L1)-mediated induction of immune tolerance has been vigorously investigated in autoimmunity and anti-tumor immunity. However, details of the mechanism by which PD-L1 is induced in CD4+ T cells are unknown. Here, we revealed the potential function of Klf1 and Egr2-mediated induction of PD-L1 in CD4+ T cells. We focused on the molecules specifically expressed in CD4+CD25-LAG3+ regulatory T cells (LAG3+ ...[more]

PMID: 29728568

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Project description:The Krüppel-like factors, KLF1 and KLF2, positively regulate embryonic β-globin expression, and have additional overlapping roles in embryonic (primitive) erythropoiesis. KLF1-/-KLF2-/- double knockout mice are anemic at embryonic day 10.5 (E10.5) and die by E11.5, in contrast to single knockouts. To investigate the combined roles of KLF1 and KLF2 in primitive erythropoiesis, expression profiling of E9.5 erythroid cells was performed. A limited number of genes had a significantly decreasing trend of expression in wild-type, KLF1-/- and KLF1-/-KLF2-/-. Among these, c-myc emerged as a central node in the most significant gene network. c-myc expression is synergistically regulated by KLF1 and KLF2, and both factors bind the c-myc promoters. To characterize the role of c-myc in primitive erythropoiesis, ablation was performed specifically in mouse embryonic proerythroblast cells. After E9.5, these embryos exhibit an arrest in the normal expansion of circulating red cells and develop anemia analogous to KLF1-/-KLF2-/-. In the absence of c-myc, circulating erythroid cells do not show the normal increase in α- and β-like globin expression, but interestingly, have accelerated erythroid maturation, between E9.5 and E11.5. This study reveals a novel regulatory network by which KLF1 and KLF2 regulate c-myc, to control the primitive erythropoietic program. Timed-pregnant KLF1+/-, KLF1+/- KLF2+/- females were anesthetized and sacrificed. E9.5 yolk sacs were dissected from the embryo, cryoprotected in 20% sucrose in PBS and frozen in OCT media. A small portion of the embryo tail was used for PCR genotyping. Eight micron frozen yolk sac sections were obtained and laser capture microdissection (LCM) was used to isolate primitive erythroid precursors. For each biological replicate, 2 to 4 yolk sacs from 2 different litters were used. Total RNA was isolated from 8 different wild-type, 3 KLF1-/-, 3 KLF1-/- KLF2-/- erythroid samples and hybridized to Affymetrix 430 A 2.0 microarrays.

Dataset Information

Expression profiling of mouse CD4+ T cells following Klf1 retrovirus vector transfection

Publications

Egr2-independent, Klf1-mediated induction of PD-L1 in CD4<sup>+</sup> T cells.

Similar Datasets

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