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. 2019 Nov 25;10:5351. doi: 10.1038/s41467-019-13259-2

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — HOXB-AS3 regulates rRNA transcription via guiding the EBP1 protein to the rDNA locus. a Relative pre-rRNA expression in OCI-AML3 cells treated with scramble or anti-HOXB-AS3 gapmers. b Occupancy of ribosomal DNA promoter repeat sequence (28S rDNA) by POLR1A in scramble versus HOXB-AS3 KD OCI-AML3 cells. c Sucrose gradient-based isolation and quantification (via ultraviolet radiation absorbance) of ribosomes and polysomes in scramble versus HOXB-AS3 KD OCI-AML3 cells. d Effect of HOXB-AS3 depletion on de novo protein synthesis in OCI-AML3 cells,The flow cytometry graph from one representative experiment is shown. e Comparison of the mean fluorescence intensity of the Alexa Fluor 488-positive OCI-AML3 cells treated with scramble versus HOXB-AS3 KD, in de novo protein synthesis assays. Results of four independent experiments are shown. f Relative 5-ETS pre-rRNA expression in human AML patient blasts treated in vivo with scramble versus anti-HOXB-AS3 gapmers. Results of three scramble and four HOXB-AS3 KD-treated mice are depicted in aggregate. g, h Effect of HOXB-AS3 KD on de novo protein synthesis in in vivo treated human AML blasts. i Relative pre-rRNA expression in K-562 cells transfected with empty vector (control) or HOXB-AS3 overexpressing vectors. j Occupancy of 28S rDNA by POLR1A in K-562 cells transfected with control or HOXB-AS3 overexpressing vectors. k Relative luciferase-reporter (luc/pRL-TK) activity in K-562 cells transfected with empty control (pIRES-Luc) or rDNA-promoter-containing (pHrD-IRES-Luc) luciferase-reporter vectors, and control or HOXB-AS3 overexpressing vectors. l Comparison of the mean fluorescence intensity of the Alexa Fluor 488-positive K-562 cells, transfected with control or HOXB-AS3 overexpressing vectors in de novo protein synthesis assays. Results of three independent experiments are shown. m Occupancy of 28S rDNA by EBP1 in scramble versus HOXB-AS3 KD OCI-AML3 cells. n Enrichment of 28S rDNA or the ACTB promoter region in RAP-DNA-based isolation of chromatin with control or HOXB-AS3-hybridizing biotinylated probes. P values were calculated using paired two-sided t-tests. In the figures, heights of boxplots indicate mean values with standard deviation. Error bars indicate highest and lowest values in each population. Source data are provided as a Source Data file.