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. 2021 Jun 15;35(10):2784–2798. doi: 10.1038/s41375-021-01307-0

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© The Author(s), under exclusive licence to Springer Nature Limited 2021

This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.

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Fig. 1 — A LONA lncRNA is overexpressed in NPM1 mutated AML patients. Relative expression of the LONA lncRNA between NPM1 wild type (NPM1wt) and NPM1 mutated (NPM1c + ) AML patients. The LONA RNA level of 174 patients (80 AML patients NPM1wt and 94 AML patients NPM1c + ) were evaluated by large scale quantitative real-time PCR (Fluidigm 96.96 Dynamic Array) [36]. Results were normalized to the expression of RNAseP, 5 S rRNA, and MLN51 and presented as relative expression [2^-ddCt] ± Standard Deviation (SD). ***P < 0.001. B LONA is mainly cytoplasmic in NPM1wt OCI-AML2, NB4, and HL60 cell lines, and C LONA is nuclear in NPM1c + OCI-AML3 cells. RNA level of LONA lncRNA in nuclear and cytoplasmic fractions were evaluated by RT-qPCR after cell fractionation. Results are presented as percentage of RNA level. GAPDH was used as a positive control for cytoplasmic RNA fraction and SNORD44 was used as a positive control for nuclear RNA fraction. Bar graph represents the average from four independent experiments ± Standard Deviation (SD). **P < 0.005, ***P < 0.001. Unpaired t-test. D Cytosolic localization of NPM1c + in the OCI-AML2 stably expressing Flag-GFP tagged NPM1c + (left). Cytospin cells were fixed and stained with DAPI. GFP fluorescence was examined with a confocal microscope. The mutant form of NPM1 leads to LONA nuclear delocalization (right). LONA RNA level in nuclear and cytoplasmic fractions were evaluated by RT-qPCR after cell fractionation of OCI-AML2 + NPM1c + cell line. Results are presented as percentage of RNA level. GAPDH was used as a positive control for cytoplasmic RNA fraction and SNORD44 was used as a positive control for nuclear RNA fraction. The bar graph represents the average from four independent experiments ± Standard Deviation (SD). **P < 0.005, ***P < 0.001. Unpaired t-test. E LONA is cytoplasmic in NPM1wt AML patients and mainly nuclear in NPM1c + AML patients. RNA level of LONA lncRNA in nuclear and cytoplasmic fractions were evaluated by RT-qPCR after cell fractionation. Results are presented as percentage of RNA level. HPRT was used as a positive control for cytoplasmic RNA fraction and SNORD44 was used as a positive control for nuclear RNA fraction. Bar graph represents the average from three independent experiments ± Standard Deviation (SD). **P < 0.005, ***P < 0.001. Unpaired t-test.