Gfi1 controls the transition from stage 2 to stage 3 during thymic iNKT cell development.

<p><b>(A)</b> The results of the quantitative RT-PCR analysis of the <i>Gfi1</i> mRNA expression in thymic iNKT cells. The developmental stage of iNKT cells was defined by the expression of CD24, CD44 and NK1.1. The results are presented relative to the mRNA expression of <i>18s</i> ribosomal RNA with the standard deviation (n = 3). <b>(B, C, D</b> and <b>E)</b> The results of the flow cytometric analyses of thymic iNKT cells in <i>Gfi1</i> KO mice. A typical CD3εand α-GalCer-loaded CD1d tetramers pattern is shown <b>(B)</b>. The typical pattern of CD24 and α-GalCer-loaded CD1d tetramers (left) and the absolute numbers with the standard deviation of stage 0 (CD24^pos) iNKT cells (right, n = 3) is indicated <b>(C)</b>. The CD1d-tetramer^pos thymocytes were enriched using a magnetic cell sorter and stained with anti-CD24 and anti-TCRVβ8 mAbs. The typical staining pattern of CD24 and α-GalCer-loaded CD1d tetramers (left), TCRVβ8 gated on CD24^high, α-GalCer-loaded CD1d tetramer^pos cells (middle), and the absolute numbers with the standard deviation of Vβ8^pos, CD24^high, and α-GalCer-loaded CD1d tetramer^pos iNKT cells (right, n = 3) are indicated <b>(D)</b>. The typical pattern of CD24 and NK1.1 gated on α-GalCer-loaded CD1d tetramer-positive iNKT cells (left) and the absolute numbers with the standard deviation of stage 1–2 (CD24^lowNK1.1^neg) iNKT cells and stage 3 (CD24^lowNK1.1^pos) iNKT cells (right, n = 3) is indicated <b>(E)</b>. <b>(F)</b> The typical pattern of CD44 and NK1.1 gated on α-GalCer-loaded CD1d tetramer-positive iNKT cells (left) and the absolute numbers with the standard deviation of stage 1 (CD44^lowNK1.1^neg), stage 2 (CD44^highNK1.1^neg) and stage 3 (CD44^highNK1.1^pos) iNKT cells (right, n = 3) is indicated. <b>(G)</b> The results of the quantitative RT-PCR analysis of the <i>Il17rb</i> mRNA expression in WT and <i>Gfi1</i>-deficient thymic iNKT cells. The results are presented relative to the mRNA expression of <i>18s</i> ribosomal RNA with the standard deviation (n = 3). <b>(H)</b> The absolute numbers of CD4^posCD8^pos, CD4^posCD8^neg, CD4^negCD8^pos and CD4^negCD8^neg iNKT cells in WT and <i>Gfi1</i> KO mice (n = 5 per group). NS; not significant, *P<0.05, **P<0.01 (Student’s <i>t</i>-test).