NFATc1 deletion in T lymphocytes inhibits the allergic trait in a murine model of
asthma
Sonja Koch, PhD1, Sarah Reppert, PhD1, Susetta Finotto. PhD1
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Laboratory of Cellular and Molecular Lung Immunology, Department of Molecular
Pneumology, Friedrich-Alexander-Universität Erlangen-Nürnberg, Germany
Running Title: NFATc1 in T cells induces allergy
Corresponding Author:
Prof. Susetta Finotto, PhD
Laboratories of Cellular and Molecular Lung Immunology
Institute of Molecular Pneumology,
Friedrich-Alexander-Universität Erlangen-Nürnberg
Hartmannstraße 14
91052 Erlangen, Germany
Phone: 0049-9131-8535883
Fax: 0049-9131-8535977
Email: susetta.finotto@uk-erlangen.de
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Abbreviations
AHR: airway hyper responsiveness
ATF: activating transcription factor
BAL: bronchoalveolar lavage
BALF: bronchoalveolar lavage fluid
BATF: Basic leucin zipper transcription factor ATF-like
CCL17: chemokine ligand 17
CCR: chemokine receptor
GATA3: GATA-binding protein 3
IFNγ: interferon gamma
Ig: Immunoglobulin
IL: interleukin
i.n. intranasally
i.p.: intraperitoneally
NFATc1: Nuclear factor of activated T cells c1
OVA: ovalbumin
PAS: Periodic Acid-Shiff
RORγT: RAR-related orphan receptor gamma T
TARC: thymus- and activation-regulated chemokine
T-bet: T-box expressed in T cells
Th: T helper cell
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Table S1 List of the used mouse qPCR primers
Gene
Gata3
Maf
Batf
Il-4
Il-13
Il-10
Il-21
Il-2
CD40l
Nfatc1ex3
Nfatc1 A
Fasl
Cd4
Hprt
Primer pairs 5´-3´
mouse
GTCATCCCTGAGCCACATCT
TAGAAGGGGTCGGAGGAACT
AGCAGTTGGTGACCATGTCG
TGGAGATCTCCTGCTTGAGG
GTTCTGTTTCTCCAGGTCC
GAAGAATCGCATCGCTGC
TCAACCCCCAGCTAGTTGTC
AAATATGCGAAGCACCTTGG
CAGCTCCCTGGTTCTCTCAC
CCACACTCCATACCATGCTG
CCAAGCCTTATCGGAAATGA
TTTTCACAGGGGAGAAATCG
CAG GAGGGGAGGAAAGAAAC
GGGAATCTTCTCGGATCCTC
AACCTGAAACTCCCCAGGAT
TCATCGAATTGGCACTCAAA
TTGCAGCACAGTTGTAAGC
TGAATGGGCGTTGACTCGAA
TGCCTTTTGCGAGCAGTATCT
CAGGCAAGGATGGGCTCATAT
ACCTGTGCAAGCCAAATTCC
AGAGTTACCATTGGCAGGAAG
GGCTCCTCCAGGGTCAGTTT
GGGGTTGGCTATTTGCTTTTC
AGGAAGTGAACCTGGTGGTG
CTCCTGCTTCAGGGTCAGTC
GCCCCAAAATGGTTAGGTT
TTGCGCTCATCTTAGGCTTT
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Figure S1 Analysis of transcription factors and cytokines under Th2 skewing conditions
(A) Schematic overview of the Nfatc1 gene and isoforms with primer binding sites (red). (BE) qPCR analysis of Nfatc1 exon 3 mRNA (B and C) and Nfatc1 isoform A mRNA (D and E)
in in vitro Th2 skewed splenic T cells isolated from NFATc1fl/fl control mice. (F-J) Total RNA
was extracted from the differentiated Th2 cells and qPCR was performed after 48h and 5 days
of culture for Gata3 (F), Maf (G and H) and Batf (I and J) in NFATc1xCreCD4 and
NFATc1fl/fl control mice. (K-N) qPCR analysis of Il-4 mRNA (K) and Il-13 mRNA (L) and Il10 mRNA (M) of the Th2 differentiated splenic cells 48h after culture and Il-10 mRNA
expression after 5 days of culture (N). n = 4 mice per group. Statistical significances in this
figure were evaluated with a Students t test. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001. Data are
mean ± s.e.m.
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