American Society of Hematology

Figure 6.

$circMYBL2 interacts with PTBP1 to affect FLT3-ITD AML proliferation through promoting FLT3 kinase translation. (A-B) Western blot showing downregulation of STAT5 signaling activation upon PTBP1 knockdown in MOLM-13 and MV4-11 cells and in FLT3-ITD AML patient samples. (C) Schematic of the circMYBL2 overexpression vector. (D) Western blot showing upregulation of FLT3 protein expression upon overexpression of either circMYBL2 or PTBP1 in MV4-11 and HL60 cells. (E) Polysomes in cytoplasmic extracts of MV4-11 cells with PTBP1 knockdown or control treatment were fractionated through sucrose gradients, and the relative levels of FLT3 mRNA in the gradient fractions were analyzed by qRT-PCR. (F) Western blot showing upregulation of FLT3 protein expression upon overexpression of either circMYBL2 or PTBP1 in 293t cells. (G) Western blot measuring FLT3 protein expression in 293t cells with overexpression of circMYBL2 in the presence or absence of si-PTBP1. (H) RIP of PTBP1 using anti-PTBP1 or anti-IgG antibodies in 293t cells in the presence or absence of circMYBL2 overexpression. (I) Schematic depicting the dominant function of circMYBL2 in facilitating the translational efficiency of FLT3 kinase by enhancing the binding of PTBP1 to FLT3 mRNA, specifically promoting FLT3-ITD AML progression. CMV, cytomegalovirus.$

circMYBL2 interacts with PTBP1 to affect FLT3-ITD AML proliferation through promoting FLT3 kinase translation. (A-B) Western blot showing downregulation of STAT5 signaling activation upon PTBP1 knockdown in MOLM-13 and MV4-11 cells and in FLT3-ITD AML patient samples. (C) Schematic of the circMYBL2 overexpression vector. (D) Western blot showing upregulation of FLT3 protein expression upon overexpression of either circMYBL2 or PTBP1 in MV4-11 and HL60 cells. (E) Polysomes in cytoplasmic extracts of MV4-11 cells with PTBP1 knockdown or control treatment were fractionated through sucrose gradients, and the relative levels of FLT3 mRNA in the gradient fractions were analyzed by qRT-PCR. (F) Western blot showing upregulation of FLT3 protein expression upon overexpression of either circMYBL2 or PTBP1 in 293t cells. (G) Western blot measuring FLT3 protein expression in 293t cells with overexpression of circMYBL2 in the presence or absence of si-PTBP1. (H) RIP of PTBP1 using anti-PTBP1 or anti-IgG antibodies in 293t cells in the presence or absence of circMYBL2 overexpression. (I) Schematic depicting the dominant function of circMYBL2 in facilitating the translational efficiency of FLT3 kinase by enhancing the binding of PTBP1 to FLT3 mRNA, specifically promoting FLT3-ITD AML progression. CMV, cytomegalovirus.

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