Fig. 5
HOTAIRM1 promotes EGR1 degradation through MDM2-mediated ubiquitination. a Western blot analysis of EGR1 levels in the different transfection groups of OCI-AML3 and OCI-AML2 cells. b Relative EGR1 mRNA expression in HOTAIRM1-silenced and HOTAIRM1-overexpressing cells. c EGR1 protein expression in transfected OCI-AML3 and OCI-AML2 cells after treatment with MG132. d EGR1 expression in transfected OCI-AML3 and OCI-AML2 cells treated with CHX. e Western blot analysis of endogenous EGR1 ubiquitination in transfected OCI-AML3 and OCI-AML2 cells. f IP assays were used to investigate the binding of MDM2 to EGR1 in OCI-AML3 cells. g The level of EGR1 and MDM2 were examined by western blotting in OCI-AML3 and OCI-AML2 cells transduced with shMDM2 or transfected with the MDM2-Flag plasmid. h Western blot analysis of EGR1 ubiquitination in OCI-AML2 cells transfected with the MDM2-Flag plasmid. i The interactions between EGR1 or MDM2 and HOTAIRM1 were validated by RNA pulldown and subsequent western blotting. j The binding of MDM2 to EGR1 in HOTAIRM1-silenced and HOTAIRM1-overexpressing cells was investigated by IP assays. 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