Article Title: The homeoprotein Dlx5 drives murine T-cell lymphomagenesis by directly transactivating Notch and upregulating Akt signaling
Figure Lengend Snippet: Lck-Dlx5 lymphomas exhibit a molecular signature reminiscent of human T-ALL ( A ) RNA microarray heatmap demonstrating that compared to thymocytes from three WT mice, primary T-cell lymphomas from three Lck-Dlx5 mice exhibit upregulation ( Red ) of genes involved in Notch signaling, cell cycle progression, chromatin modification, calcium homeostasis, glucose, fatty acid and nucleotide metabolism, as well as classical Dlx5 target genes. Down-regulated genes ( Green ) are implicated in pro-apoptotic events ( Tnf , Fas , Xaf1 , INFb signaling-related genes), tumor suppression, and DNA damage repair. ( B ) compared to T-cell lymphomas from Lck-MyrAkt2 mice, Lck-Dlx5 lymphomas show upregulation of Notch1/3 and downregulation of Lats2 , Appl2 , Jak3 , Tet2 , and Tnik . Note that in the interests of journal space, the heatmap shown here includes only one sample from each group. The full heatmaps for all three samples examined per group can be found in Supplementary Figure 2C. ( C ) real-time PCR on three T-cell samples from WT mice (samples 1–3), 11 primary lymphomas from Lck-Dlx5 mice (samples 4-14: F86-785, -801, -793, -1149; F63-0, -1263, -1210; F47-0, -1247, -918; and F84-1063), and 7 lymphomas from Lck-MyrAkt2 mice (samples 15-21: F72-918, -2811, -3148, -3154; and F420-577, -1174, -1073), confirming unique upregulation of Notch signaling in Dlx5-driven tumors. Data = mean ± SEM. ( D ) immunoblot showing overexpression of Notch1, NIC1, Notch3, Hes1 and Myc in lymphoma lines (F63-0, F47-0, F47-918) from Lck-Dlx5 mice. Loss of Pten expression and corresponding high levels of phospho-Akt in Lck-Dlx5 -derived tumor cells are also shown, as are cyclin D1 levels. Dlx5 proteins have high molecular weight (a, b) and low molecular weight (c) forms. Notch proteins have full length ( F ) and cleaved (C) forms. *, non-specific bands. ( E ) H E and immunohistochemical staining of lymphoma invading lung of Lck-Dlx5 mouse. Note strong staining for Notch1, Notch3, Hes1, Myc and phospho-Akt in lymphoma. (F) summary of real-time PCR analysis of RNA from 15 pediatric T-ALL specimens showing expression levels of DLX5 , NOTCH1 and NOTCH3 .
Article Snippet: Myc-Tag antibody, as well as IgG control, were used to immunoprecipitate Dlx5/chromatin complexes, and captured DNA fragments were analyzed by Illumina GA II to obtain whole-genome datasets (available at GEO #GSE83778).
Techniques: Microarray, Mouse Assay, Modification, Real-time Polymerase Chain Reaction, Over Expression, Expressing, Derivative Assay, Molecular Weight, Immunohistochemistry, Staining