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. 2019 May 8;10:1016. doi: 10.3389/fimmu.2019.01016

Figure 3.

Figure 3

Hematopoietic-specific deletion of Foxo1 downregulates cell-cycle repressors of NK cells. (A) q-RT-PCR analysis of direct target genes of Foxo1 responsible for cell cycle in sorted splenic NK cells (CD3−CD19−NKp46+, purity >95%) from control and Foxo1Δ/Δ mice. Each dot represents one mouse. 5 littermates were included for p21cip1; 4 littermates were included for p27kip1, Gadd45a, p130, cyclinD2, and Ccng2; and 2 replicates were included for cyclinD1 (Error bars indicate SD; unpaired Student's t-test with generalized linear models; *p < 0.05, **p < 0.01, and ***p < 0.001, control vs. Foxo1Δ/Δ mice). (B) Schematic structure of indicated mouse cell-cycle repressors with putative Foxo1 binding sites (Top panels). ChIP assay of Foxo1 binding to the promoter region of indicated cell-cycle repressors in sorted splenic NK cells (CD3−CD19−NKp46+) from wild type mice (Bottom panels). The precipitated DNA by Foxo1 was performed with PCR. Representative PCR gel pictures from 1 of 2 replicates are shown.