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Mediators of Inflammation
Volume 2017, Article ID 5420718, 15 pages
Research Article

Key Role of STAT4 Deficiency in the Hematopoietic Compartment in Insulin Resistance and Adipose Tissue Inflammation

1Department of Physiological Sciences, Strelitz Diabetes Center, Eastern Virginia Medical School, Norfolk, VA, USA
2Department of Internal Medicine, Strelitz Diabetes Center, Eastern Virginia Medical School, Norfolk, VA, USA
3Department of Pediatrics, Indiana University School of Medicine, Indianapolis, IN, USA

Correspondence should be addressed to Anca D. Dobrian; ude.smve@daairbod

Received 12 November 2016; Revised 22 December 2016; Accepted 25 December 2016; Published 16 March 2017

Academic Editor: Dah-Yuu Lu

Copyright © 2017 Anca D. Dobrian et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Supplementary Material

Supplemental Figure 1 illustrates the general experimental approach. Bone marrow transplant is shown in Protocol 1. Males, 8 weeks old STAT4-/- mice and Stat4+/+ littermate controls were sub-lethally irradiated with two 6Gy doses applied 4 hours apart and reconstituted via tail vein injections with bone marrow cells (BMC) isolated from either STAT4+/+ mice or STAT4-/- homozygotes. STAT4-/- recipients received BMCs from STAT4+/+ mice and STAT4+/+ recipients received BMCs from STAT4-/- mice. Mice were allowed to fully recover for 4 weeks and were subsequently placed on 60%kcal fat diet (HFD) for 15 weeks. Protocol 2 shows adoptive transfer in Rag1-/- mice. Male, 12 weeks old Rag1-/- mice were adoptively transferred via tail vein injection with one of the following: STAT4+/+CD4+, STAT4-/- CD4+, STAT4+/+CD8+ or STAT4-/-CD8+ cells isolated from spleens of STAT4+/+ or STAT4-/- donor mice. All mice were placed on 60%kcal fat for 15 weeks. Supplemental Figure 2 shows tissue chimerism following bone marrow transplant. Representative agarose gels show the spleen, bone marrow and blood chimerism following bone marrow transplantation. DNA was isolated from each of the tissues and amplified using specific primers to detect the deletion of the STAT4 gene. In both recipient groups the chimerism with donor cells was close to 100%. Spleens of recipient mice in both groups showed a similar >85% chimerism. Histogram shows densitometry results for % donor genotype out of total signal.

  1. Supplementary Material