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. 2015 Jun 1;212(6):953–970. doi: 10.1084/jem.20150002

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© 2015 Thompson et al.

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Figure 6. — Dyrk1a-deficient quiescent DP thymocytes and small pre–B cells fail to repress E2F target gene transcription. (A and B) Up- and down-regulated transcripts were identified by RNA-sequencing of FACS-purified quiescent DP thymocytes (CD4⁺/CD8⁺/DNA content^2N/Pyronin Y^low), cycling DP thymocytes (CD4⁺/CD8⁺/DNA content^>2N/Pyronin Y^high), small pre–B cells (IgM⁻/B220⁺/CD43⁻/FSC^low), large pre–B cells (IgM⁻/B220⁺/CD43^low/FSC^high), and granulocytes (Gr-1^high/Mac-1/CD11b^high) from Dyrk1a^f/f Mx1-Cre⁻ (Control) and Dyrk1a^f/f Mx1-Cre⁺ (CKO) mice. Venn diagrams depict shared up-regulated (A) and down-regulated (B) transcripts identified by RNA-sequencing in the three quiescent cell populations. (C–F) mRNA expression of E2F target genes was assessed by qRT-PCR in FACS-purified small pre–B cells (C), quiescent DP thymocytes (D), large pre–B cells (E), and cycling DP thymocytes (F) from Control and CKO mice 2 wk after pI:pC treatment. Transcript levels were normalized to Actb expression. PCRs were performed using 3 independently sorted pairs of samples (each with 1 mouse per genotype) for pre–B cells, and pooled samples from 3 mice per genotype for thymocytes. Error bars depict SD of triplicate wells for representative samples. *, P < 0.05; **, P < 0.01; ***, P < 0.001; NS, not significant (CKO vs. Control).