Figure 6.
miR-155 represses multiple targets in mouse FLT3-ITD LK and human FLT3-ITD+AML cells, including the IFN gene regulator Cebpb. (A) Expression level of miR-155 targets in the LKS compartment of FLT3-ITD or FLT3-ITD 155−/− mice by RNA sequencing. (B) Expression level of miR-155 targets in the TCGA human AML data set. (C) Western blot of SHIP1, PU.1, and CEBPB in LK-sorted BM of FLT3-ITD or FLT3-ITD 155−/− mice; replicates represent individual mice. (D) qRT-PCR analysis of miR-155 targets Cebpb, Ship1, and Pu.1 in Molm14 and MV4-11 cells infected with either EV or 155-CR1 constructs. Expression normalized to L32, with the EV-infected cell average set to a relative expression value of 1. n = 3 biological replicates for each condition. (E-F) qRT-PCR analysis of FLT3-ITD LK cells infected with either the pMIGII-EV or pMIGII-Cebpb vector. Expression normalized to L32 with pMIGII-EV–infected cell average set to a relative expression value of 1. n = 3 biological replicates for each condition. (G) Western blot of phospho-AKT (p-AKT) and total AKT of LK cells from FLT3-ITD and FLT3-ITD 155−/− mice; replicates represent individual mice. Data represented as mean ± SEM. *P ≤ .05; **P ≤ .01; ***P ≤ .001.

miR-155 represses multiple targets in mouse FLT3-ITD LK and human FLT3-ITD+AML cells, including the IFN gene regulator Cebpb. (A) Expression level of miR-155 targets in the LKS compartment of FLT3-ITD or FLT3-ITD 155−/− mice by RNA sequencing. (B) Expression level of miR-155 targets in the TCGA human AML data set. (C) Western blot of SHIP1, PU.1, and CEBPB in LK-sorted BM of FLT3-ITD or FLT3-ITD 155−/− mice; replicates represent individual mice. (D) qRT-PCR analysis of miR-155 targets Cebpb, Ship1, and Pu.1 in Molm14 and MV4-11 cells infected with either EV or 155-CR1 constructs. Expression normalized to L32, with the EV-infected cell average set to a relative expression value of 1. n = 3 biological replicates for each condition. (E-F) qRT-PCR analysis of FLT3-ITD LK cells infected with either the pMIGII-EV or pMIGII-Cebpb vector. Expression normalized to L32 with pMIGII-EV–infected cell average set to a relative expression value of 1. n = 3 biological replicates for each condition. (G) Western blot of phospho-AKT (p-AKT) and total AKT of LK cells from FLT3-ITD and FLT3-ITD 155−/− mice; replicates represent individual mice. Data represented as mean ± SEM. *P ≤ .05; **P ≤ .01; ***P ≤ .001.

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