Fig. 8: Adiponectin facilitates T-cell selection and development within the TNC complexes.

a WT and AKO mice were sacrificed at the age of 6–7 weeks. Thymus (top) and TNC complexes (bottom) were prepared to analyze the percentage of CD3⁺ cell population by flow cytometry. b Four-week-old WT mice were subjected to vehicle treatment or adoptive transfer with EGFP⁺ cells from thymus of Adn-Cre/ROSA^mT/mG mice (30000 cells/mouse via tail vein) as described in the Methods. At 21 days after the injection, thymus was collected and subjected to flow cytometry analyses for measuring the percentage contents of CD3⁺ (top) or CD4⁺CD8⁻ (bottom) cells. c WT and AKO mice were sacrificed as in (a) to prepare thymic cell suspensions. Flow cytometry was performed to analyze the total amount of CD326⁺β5t⁺ TNC complexes (top), TNC complexes with high expression levels of propidium iodide [PI] (middle), or eCD45⁺ cells attached outside of the TNC complexes (bottom). d WT and AKO mice were sacrificed as in (a) to prepare the thymic cell suspensions enriched with TNC complexes. Inverted microscope was used to visualize and manually count the number of thymocytes within each TNC complex (top). Immunofluorescence staining was performed to detect CD3 (middle) and β5t (bottom) signals in TNC complexes of WT and AKO mice. The slides were counterstained with DAPI. Data are presented as mean ± SEM. *, P < 0.05 and **, P < 0.01 vs corresponding controls (n = 6 biologically independent samples from different animals of independent experiment).