Figure 3

YY1^lo NKT cells are found in lymphoid and nonlymphoid tissues and share a common transcription factor expression pattern. Lymphocytes were isolated from the spleen, visceral adipose, lung, and liver of 12 week old, C57BL/6J (WT) mice and analyzed by FACS with the indicated antibodies. Expression of PLZF and YY1 in the NKT cells (CD45.2⁺, MHCII⁻, CD3⁺ CD1dtet⁺ CD24⁻) was determined. (a) Representative FACs plots and (b) cumulative data comparing the frequency of the YY1/PLZF subpopulations between tissues (c–e). Intracellular staining for (c) Tbet and PLZF or (d) RORγt and PLZF in total spleen NKT cells (left), YY1^hi NKT cells (middle), and YY1^lo NKT cells (right). (e) Distribution of spleen YY1^lo and YY1^hi NKT cells in the NKT1, NKT2, and NKT17 subpopulations as defined in (c,d). (f,g) ThPOK expression was compared between spleen YY1^lo and YY1^hi NKT cells by FACs. A representative histogram is shown in (f) and cumulative data are summarized in (g). (h,i)Expression of CD44 and NK1.1 was compared between the YY1/PLZF subpopulations. Data in (a,c,d,f,h) are representative of at least 4 independent experiments. Each dot in (b,e,g,i) represents an individual mouse, for (b) N ≥ 5 for each tissue and for (e,g) N = 4, for (i) N = 6. The horizontal lines indicate the mean (±s.e.m.). *P < 0.05 **P < 0.01, ***P < 0.001 determined by one-way ANOVA (b,i) or Mann-Whtiney U Test (e,g).