Journal: Nature immunology

Article Title: TGF-β is responsible for NK cell immaturity during ontogeny and increased susceptibility to infection during mouse infancy

doi: 10.1038/ni.2388

Figure Lengend Snippet: ( a – d ) CD11c dnR and wild type mice were injected ( i.p. ) twice a day with BrdU for 3 days. On day 3, mice were sacrificed and frequency of cycling cells was determined in the bone marrow. ( a – b ) FACS plots show the distribution of BrdU staining among total mNK cells ( a ) and gated mNK cells at stages D, E, and F ( b ). ( c ) Graph shows the frequency of cycling cells in pNK, iNK, and mNK cells from CD11c dnR (black circle) versus wild type (white circle) mice. ( d ) Graph shows the frequency of cycling cells in mNK cells at stages D, E, and F from CD11c dnR (black circle) versus wild type (white circle) mice. Data in a , d are representative of three independent experiments with n = 2 mice per experiment and results in c , d show all 6 individual mice. ( e – f ) mNK cells at stages D, E, and F were sorted from the bone marrow. mRNA was isolated and cDNA was subjected to pathway-specific qPCR for analysis of cell cycle genes ( e ) or SYBR Green qPCR for analysis of transcription factors T-bet, GATA-3, and IRF-2 ( f ). Data were analyzed using Global Pattern Recognition analytical software ( e ) or 2-33C3 method ( f ) and results were expressed as fold of change in CD11c dnR versus wild type samples. Data in e , f are representative of three independent cell sorting with samples pooled from n = 12 CD11c dnR and 25 WT mice.

Article Snippet: For cell cycle genes, we used a customized cell cycle qPCR array according to the manufacturer’s instructions (Lonza), and data were analyzed using Global Pattern Recognition analytical software (Lonza).

Techniques: Injection, BrdU Staining, Isolation, SYBR Green Assay, Software, FACS