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FLK1+ cells were isolated on Myltenyi magnetic columns.

Human peripheral blood mononuclear cells were isolated on a Ficoll-Pacque gradient and total RNA was prepared by using an RNA Purification kit (QIAGEN, Valencia, CA, USA).

CD4+CD25+ and CD4+CD25− MLN cells were isolated on day 23 after arthritis induction and cultured with HSP60 peptide 180-188 or media.

Cells were isolated on ice, after red blood cell lysis, cells were stained for Lin−c-kit+CD41+ using the same antibody clones than mentioned above, except for c-kit which was conjugated to PeCy7 instead of APC.

Tregs and Tconv cells were isolated on a BD FACSAria II high speed cell sorter following the CD4+CD45RA+ pre-enrichment (BD Biosciences, San Jose, CA) with the following antibodies: CD4-PerCP (clone SK3), CD127-PE (hIL-7R-M21), and CD25-APC (2A3) as previously described [24], with the addition of CD45RA-PE-Cy7 (HI100) to select the naive Treg subset.

Splenic CD8+CD25+ T cells were isolated on day 6 after viral infection.

The region of interest of the monocyte population, comprising over 3,000 cells, was isolated on morphological criteria of cell size and granularity.

7 days after T-cell transfer, spleen cells were isolated to examine IL-7Rα expression on donor CD4+ T cells.

Fluorescent cells were isolated based on forward and side scatter characteristics and fluorescence expression.

T cells were isolated based on FoxP3GFP expression and transferred to LDLr−/−, Rag−/− mice to establish a role for B6.SLE effector T cells (Teff) in atherosclerosis.

Ovine oviduct epithelial cells were isolated based on the protocol by R. Ian Freshney and Mary G. Freshney [ 24], and all the procedures were performed under sterile conditions.