Figure 1. Id2 and Id3 expression in aTregs.

(A) qPCR showing Id2 and Id3 mRNA expression in sorted CD4⁺ CD25⁺ Tregs isolated from the spleen or visceral adipose tissue. Id2 or Id3 expression was normalized to HPRT. Each dot indicates cells isolated from one animal. (B) Flow cytometry histograms showing Id2-YFP or ID3-GFP expression in gated CD4⁺ CD25⁺ Tregs isolated from the spleen or visceral adipose tissue. B6 mice were used as a ‘no reporter’ control. Numbers in histograms indicate the median fluorescence intensity (MFI). Bar graphs indicate the average MFI in Tregs isolated from the indicated tissue. (C) Histograms indicating the frequency of Id2⁺ or Id3⁺ Tregs identified in the spleen (left) and their expression by flow cytometry of ST2 and KLRG1 (right). Yellow peaks indicate Id2-YFP⁺ cells and green peaks indicate Id3-GFP⁺ cells. Grey peaks indicate Id2-YFP⁻ or Id3-GFP⁻ cells, respectively. Data are representative of two independent experiments with 1-3 mice per group. P values were calculated using the student’s t-test.