Fig. 4

HOTAIRM1 interacts with EGR1 in NPM1-mutated AML cells. a The subcellular localization of HOTAIRM1 in OCI-AML3 cells was detected by FISH. Scale bar: 25 μm. b Schematic diagram of the identification of HOTAIRM1 binding proteins by RNA pulldown coupled with mass spectrometry. c The interactions between EGR1 or Ago2 and HOTAIRM1 in OCI-AML3 cells were validated by RNA pulldown and subsequent western blotting. d HOTAIRM1 RNA was measured by qRT-PCR in IgG control, EGR1, and Ago2 RIP samples. e Schematic diagram of the putative EGR1-binding sites in HOTAIRM1. f The effect of overexpression of HOTAIRM1-WT, HOTAIRM1-Mut1 and HOTAIRM1-Mut2 in HOTAIRM1-silenced OCI-AML3 cells was assessed by RIP assays. g RNA pulldown and western blotting were used to detect EGR1 expression in HOTAIRM1-silenced OCI-AML3 cells transfected with different vectors. The data are presented as the mean ± SD of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001. n.s. indicates no significant difference