Figure 5.
WTAs for tumors found in p53−/−VAV1-Tg or p53−/− mice. (A) Unsupervised hierarchical clustering analyses for T-LBL from mice of indicated genotypes and whole thymocytes from WT mice (i), and tumor cells sorted from Lym from mice of indicated genotypes and CD4+ or CD8+ splenocytes from WT mice (ii). (B) Venn diagram of genes expressed at threefold higher levels in T-LBL from p53−/−VAV1-Tg mice relative to p53−/− mice, or levels seen in Lym from p53−/−VAV1-Tg mice relative to CD4+ splenocytes. Thirty-six genes highly expressed in both of T-LBL and Lym of p53−/−VAV1-Tg mice compared with controls are listed. (C) Myc expression levels in T-LBL from p53−/− mice of (n = 2), and p53−/−VAV1-Tg mice (n = 4), and in CD4+ splenocytes from WT mice (n = 3), and Lym from p53−/−VAV1-Tg mice (n = 5). (D) Significantly enriched gene sets identified by GSEA using curated gene sets in the Molecular Signatures Database.26,27 T-LBL from p53−/−VAV1-Tg mice vs T-LBL from p53−/− mice (i), and tumor cells sorted from Lym from p53−/−VAV1-Tg mice vs CD4+ and CD8+ splenocytes from WT mice (ii). Red lines indicate MYC-related gene sets. (E) Representative results of enrichment plots of Myc-related gene sets. T-LBL from p53−/−VAV1-Tg mice vs T-LBL from p53−/− mice (i), and tumor cells sorted from Lym from p53−/−VAV1-Tg mice vs CD4+ or CD8+ splenocytes from WT mice (ii). (F) GEPs of WT CD4+ or CD8+ splenocytes, tumor cells sorted from Lym from p53−/−VAV1-Tg mice, and cells from AITL-like tumors in RHOAG17VTet2−/− mice.28 (G) Cell-surface expression of Ccr4 and Gata3 analyzed by flow cytometry. (H) Immunohistochemical analysis for Gata3 in spleens of Lym from p53−/−VAV1-Tg mice. Original magnification ×40. Scale bars, 20 µm. The percentages at the bottom right corner indicate positivity in the tumor cells. (I) Schema showing a protocol for skewed differentiation of naive T cells sorted from splenocytes with each genotype. Cells were cultured in neutral condition without any cytokines or antibodies and Th2 condition with recombinant mouse IL-4 and anti–IFN-γ antibody, respectively. Gata3 expression levels were examined by flow cytometry 8 days after TCR stimulation.

WTAs for tumors found in p53−/−VAV1-Tg or p53−/− mice. (A) Unsupervised hierarchical clustering analyses for T-LBL from mice of indicated genotypes and whole thymocytes from WT mice (i), and tumor cells sorted from Lym from mice of indicated genotypes and CD4+ or CD8+ splenocytes from WT mice (ii). (B) Venn diagram of genes expressed at threefold higher levels in T-LBL from p53−/−VAV1-Tg mice relative to p53−/− mice, or levels seen in Lym from p53−/−VAV1-Tg mice relative to CD4+ splenocytes. Thirty-six genes highly expressed in both of T-LBL and Lym of p53−/−VAV1-Tg mice compared with controls are listed. (C) Myc expression levels in T-LBL from p53−/− mice of (n = 2), and p53−/−VAV1-Tg mice (n = 4), and in CD4+ splenocytes from WT mice (n = 3), and Lym from p53−/−VAV1-Tg mice (n = 5). (D) Significantly enriched gene sets identified by GSEA using curated gene sets in the Molecular Signatures Database.26,27  T-LBL from p53−/−VAV1-Tg mice vs T-LBL from p53−/− mice (i), and tumor cells sorted from Lym from p53−/−VAV1-Tg mice vs CD4+ and CD8+ splenocytes from WT mice (ii). Red lines indicate MYC-related gene sets. (E) Representative results of enrichment plots of Myc-related gene sets. T-LBL from p53−/−VAV1-Tg mice vs T-LBL from p53−/− mice (i), and tumor cells sorted from Lym from p53−/−VAV1-Tg mice vs CD4+ or CD8+ splenocytes from WT mice (ii). (F) GEPs of WT CD4+ or CD8+ splenocytes, tumor cells sorted from Lym from p53−/−VAV1-Tg mice, and cells from AITL-like tumors in RHOAG17VTet2−/− mice.28  (G) Cell-surface expression of Ccr4 and Gata3 analyzed by flow cytometry. (H) Immunohistochemical analysis for Gata3 in spleens of Lym from p53−/−VAV1-Tg mice. Original magnification ×40. Scale bars, 20 µm. The percentages at the bottom right corner indicate positivity in the tumor cells. (I) Schema showing a protocol for skewed differentiation of naive T cells sorted from splenocytes with each genotype. Cells were cultured in neutral condition without any cytokines or antibodies and Th2 condition with recombinant mouse IL-4 and anti–IFN-γ antibody, respectively. Gata3 expression levels were examined by flow cytometry 8 days after TCR stimulation.

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