Fig. 2: SET Knock down impairs the colony forming unity (CFU) ability of KMT2A-R-leukemic cells.
From: SET-PP2A complex as a new therapeutic target in KMT2A (MLL) rearranged AML
A Expression of mouse SET mRNA (probe1421819_a_at) in 12 high LSC frequency KMT2A-R AMLs (KMT2A::MLLT3 and KMT2A:MLLT1) and 22 low LSC frequency KMT2A-R-AMLs (KMT2A::AFF1p, KMT2A::AF10 and KMT2A::GAS7). Meta-analyses of micro-array data from GSE13690. Unpaired t test; **p < 0.01. B Spearman correlation matrix for SET mRNA and human LSC genetic signature obtained from ref. [36]. C Spearman correlation matrix for SET mRNA and the self-renewal associated genetic signature identified in human KMT2A-R AML cells by [37]. The genes in red have a significant positive correlation with SET (Rs >0.2 and p < 0.05), while the genes in blue have a significant negative correlation with SET (Rs < −0.2 and p < 0.05). The genes marked with a cross do not correlate with SET expression. D Immunoblot for SET in K562, Kasumi1, and REH stably expressing shSCRAMBLE, RFP or shSET. Densitometry analysis was conducted by LI-COR Image Studio software. GAPDH was used as a loading control. Values are expressed as ratio between SET and GAPDH relative to untransduced cells. E qRT-PCR showing the expression of SET in eGFP-THP1, eGFP-MV411, and eGFP-SEM expressing either shScramble or shSET. Gene expression was normalized to GAPDH control and analyzed by Pfaffl equation. Values are expressed relative to shScramble controls. Data represent mean ± SD of three independent experiments. Unpaired t test **** p < 0.0001. F–K Effect of shSET on colony-forming unit ability. Data show mean ± SD of three independent experiments. 2-way Anova Tukey’s multiple comparative tests **p < 0.01; ****p < 0.0001. K–N Proliferation curve of K562 and Kasumi1 and REH, stably expressing eGFP and shScramble or shSET. GFP expression was used as quantitative reporter of cell proliferation. For each cell line, the same number of cells was plated at t0 and the GFP signal was measured by a fluorescent microplate reader at each time point. Data show mean ± SD of triplicate wells and are representative of three independent experiments. 2-Way Anova Tukey’s multiple comparison test *p < 0.05; **** p < 0.0001.
