Supplementary Material for: Meeting the Iron Needs of Young Children

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Scan of 96 well plate containing digested tissues from animals given labelled IL-7 and a serial dilution in quadruplicate of labelled murine IL-7 seen in (A).

<p>Digested spleens are in row F, wells 3 through 8, and digested thymus tissue is in row H, wells 3 through 8. Wells A–D in column 1 contains IL-7 at 10 ng in a total volume of 50 µl and the subsequent columns (2 through 12) contain doubling serial dilutions. The standard curve of recombinant CW800 labelled murine IL-7 (B) shows means± 1 standard deviation from quadruplicate wells and the calculated amount of IL-7 in individual spleens and thymuses, are seen in (C).</p

IRDye800 labelled IL-7 introduced by gavage into the stomach of a mouse pup can be detected as green fluorescence in the anterior mediastinum on the next day (A), as indicated by the white arrow.

<p>Labelled product is apparent also around the jaws of the animal and in the intestine. The pup is in a prone position on the glass surface of the Odyssey and the image is from underneath. Two examples of samples from quantitation studies are shown here where newborn pups were fed 100 ng of labelled protein, either rIL-7 (B,C) or albumin (D,E), and culled after 24 hours and their thymus (B,D) and spleen (C,E) removed and scanned.</p

Comparison of the numbers of cells in the spleen of offspring and the effect of IL-7.

<p>Comparison of the numbers of cells in the spleen of offspring and the effect of IL-7.</p

In cross fostering experiments more CD44⁺CD25⁻ and CD44⁺CD25⁺ thymocytes are found in the thymus of IL-7 knock out animals fostered onto wild type animals (⋄) than those left with their IL-7 knock out mothers (▵).

<p>Values are similar to those obtained with wild type animals maintained on wild type mothers (▴). Values which are significantly different (where p<0.05) from the knock out animals on knock out mothers are indicated by the annotation.</p

IL-7 can be detected in the milk of mice in the offspring after birth but not for the entire period of weaning.

<p>The IL-7 content of milk was assayed by sandwich ELISA and quantified against recombinant murine IL-7 standards.</p

Diagrammatic representation of an outline of the experimental procedure to follow labelled IL-7 from the gut to the lymphoid organs.

<p>Diagrammatic representation of an outline of the experimental procedure to follow labelled IL-7 from the gut to the lymphoid organs.</p

Comparison of the numbers of cells in the major populations of the thymus in offspring and the effect of IL-7.

<p>Comparison of the numbers of cells in the major populations of the thymus in offspring and the effect of IL-7.</p

Baseline characteristics (means (S.D.) or geometric means (S.D.)).

*<p>geometric means.</p

Haemoglobin response (mean & 95% CI).

<p>Haemoglobin response (mean & 95% CI).</p