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Blood, Vol. 112, Issue 7, 2836-2846, October 1, 2008
Exogenous insulin-like growth factor 1 enhances thymopoiesis predominantly through thymic epithelial cell expansion
Blood Chu et al.
112: 2836
Supplemental materials for: Chu et al
Files in this Data Supplement:
- Figure S1. Exogenous IGF-1 does not affect peripheral T-cell population size in thymectomized mice (JPG, 51.1 KB)
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Recombinant human IGF-1 was administered by continuous infusion using subcutaneously placed osmotic pumps to deliver a dose of 100 ug/day for a total of two weeks in 8–12 week old female thymectomized C57BL/6 mice. Mice were sacrificed at the end of a total course of four weeks of continuous IGF-1 treatment. Total body weights were measured (A) to confirm delivery of IGF-1. Combined spleen and lymph node data (B–D) were obtained after four weeks of IGF-1 treatment: (B) CD4+ subsets; (C) CD8+ subsets; and (D) Total peripheral TREC. Presented data are combined data from two independent experiments, with ten mice per group. *p < 0.05 between IGF-1 and diluent treated mice for body weight only.
- Figure S2. Exogenous IGF-1 does not change the distribution of cTEC or mTEC (JPG, 77.0 KB)
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Thymic sections from IGF-1 treated and diluent treated controls were analyzed for relative distribution of cortical and medullary TEC populations by laser scanning cytometry, which offers the advantage over conventional microscopic techniques of being able to visualize an entire planar section of tissue in a single automated scan. (A) Representative images of thymic sections using Hoechst 33342 DNA staining and cortical TEC-specific (Ly51) and medullary TEC specific (UEA-1) markers. The relative proportions of cortical and medullary TEC were determined by calculating the percentage of phantom contours superimposed over the Hoechst 33342+ area that are positive for the specific TEC marker. (B) Representative data from two independent experiments showing relative quantitation of TEC-specific markers in whole thymus sections, three sections per experimental group.
- Figure S3. Exogenous IGF-1 expands multiple thymocyte subsets in PSGL-1KO mice (JPG, 38.3 KB)
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PSGL-1KO mice and age-matched wild type controls were given a two-week course of IGF-1, after which major thymocyte subsets defined by CD4/CD8 staining (A) and DN subsets defined by Lineage−/CD44/CD25 staining (B) were enumerated. n = 6–11 mice per group combined from three independent experiments; *p<0.05.
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