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. 2022 Jul 13;13:4061. doi: 10.1038/s41467-022-31574-z

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© The Author(s) 2022

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. 3 — a Scatter plot indicates the correlation analysis of RNA sequencing (x-axis) and protein mass spectrometry (y-axis) data in two different cell models (Daoy and CHLA-266) upon HHIP-AS1 knockdown (using sh-HHIP-AS1#1 and sh-HHIP-AS1#2) versus control cells (sh-scr, n = 3 independent samples per condition and cell model). b Scatter plot displaying expression correlation of DYNC1I2 and HHIP-AS1 sequencing data comparing FPKM expression values in 167 MB patient samples. Samples are color coded for MB subgroups. Statistics were done by Pearson correlation. c Representative image of co-localization of HHIP-AS1 and DYNC1I2 mRNA in Daoy obtained through two-color fluorescence in situ hybridization (FISH). White frame indicates the location of the zoom out picture at the right side. Green: HHIP-AS1 lncRNA, red: DYNC1I2 mRNA, blue: DAPI, Nucleus. Scale bar: 5 µm. This experiment was repeated twice with similar results. d Enrichment of DYNC1I2 mRNA upon HHIP-AS1 raPOOL pulldown in Daoy and CHLA-266 cell lines and HHU-ATRT1 primary cells. Bar graphs are presented as the mean ± SD of three independent experiments. e Bio-Layer interferometry was used for detecting direct interaction between DYNC1I2 mRNA and HHIP-AS1. f DYNC1I2 mRNA stability upon transfection of a control (HHIP-AS1^neg) or the HHIP-AS1 interacting sequence (HHIP-AS1^bind). Calculation was done in comparison to the mRNA level at time point “0 h” in each condition. Data are presented as the mean ± SEM of five independent experiments; Student´s two-sided t-test; *p < 0.05. g Bar graph indicating the proliferation rate or viability of Daoy cells in control condition (Ctrl), upon overexpression of DYNC1I2 (DYNC1I2 OE) and upon transient HHIP-AS1-knockdown (si-HHIP-AS1) in DYNC1I2 overexpression (DYNC1I2 OE + si-HHIP-AS1). Data are represented as the mean ± SD of at least six independent experiments normalized to the control condition. Student’s two-sided t-test; ***p < 0.001; n.s. not significant. h The immunoblot shows a representative blot of DYNC1I2 protein expression in control (Ctrl) or DYNC1I2-overexpressing (DYNC1I2 OE) cells. ACTB immunoblotting was used as loading control. This experiment was done twice with similar result. Source data and exact p-values are provided as a “Source Data file”.