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. 2016 Mar 28;126(5):1664–1678. doi: 10.1172/JCI81516

Figure 5. SYTL1 functions in human AML.

Figure 5

(A) SYTL1 silencing inhibits homing of RS4;11 AML cells in NOD/SCID mice. Bone marrow samples were analyzed for the human CD45-positive fraction by flow cytometry (representative of 3 experiments). Data are compared between shSYTL1-treated (shSytl1 #4 and #5) and control shRNA (negative control) RS4;11 cells. Efficiencies of SYTL1 silencing were confirmed by RT-PCR. GAPDH, which was run on a separate gel, is shown as a loading control of cDNA. (B) SYTL1 silencing inhibits engraftment of RS4;11 AML cells in NOD/SCID mice. Bone marrow samples were analyzed for the human CD45-positive fraction by flow cytometry (n = 3). SYTL1 knockdown by shSYTL1 was confirmed by immunoblotting; blots are representative of 3 independent experiments. (A and B) Frequencies of human CD45-positive cells in bone marrow are indicated (mean ± SEM). (C) Survival curve of NOD/SCID mice transplanted with RS4;11 cells with shSYTL1 lentiviral vectors or empty vector. (D) Suppression of cell migration by SYTL1 knockdown. RS4;11 cells were transduced with lentiviral vectors bearing shSYTL1, and frequencies of cell migration were examined in the presence or absence of CXCL12. Data represent mean ± SEM of 3 independent experiments. (E) Pairwise correlations between gene expression profiles of 526 AML samples hybridized to the Affymetrix HGU 133Plus 2.0 GeneChips identified subsets with high SYTL1, HOXA9, HOXA7, and MEIS1 expression levels. The bar next to each sample indicates AML with the following genetic mutations in red: A: t(8;21), B: t(15;17), C: inv(16), D: CEBPA double mutation, E: CEBPA single mutation, F: NPM1 mutation, and G: 11q23 abnormalities/MLL fusions. Histograms next to the bar indicate expression levels of the following: 1: SYTL1, 2: HOXA9, 3: HOXA7, 4: MEIS1. Correlation coefficients were 0.33, 0.32, and 0.40 between SYTL1 and HOXA9, HOXA7, and MEIS1, respectively. An inset is enlarged to show two subsets of MLL-associated AML. I: BREhiEVI1lo, II: BREloEVI1hi. (A, B, and D) **P < 0.01, 1-way ANOVA with Dunnett’s multiple comparison test.