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. 2021 Jun 16;12:3655. doi: 10.1038/s41467-021-23969-1

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© The Author(s) 2021

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. 7 — a Scheme for two splicing isoforms of hZC3H12D. The long and short forms encode 58 and 36 kDa proteins, respectively. Both have a common N-terminal sequence, including a ZF domain (green), and the long form has a proline-rich domain (orange; top). IHC analysis of ZC3H12D in the long form (hZC58)- and short form (hZC36)-overexpressing 786-O cells (bottom). Experiments were repeated twice with similar results. b Representative confocal microscopy images of hIL1β-mRNA-FITC uptake in the hZC58 or hZC36 cell nucleus (top). Quantitative signals of hIL1β-mRNA-FITC in the hZC58 or hZC36 cell nucleus (bottom; 3D images per group: n = 38 for hZC58 and n = 50 for hZC36). c Uptake of hIL1β-mRNA-FITC into CD56⁺CD3^-NK cell nucleus 3 h after applying 50 ng/mL RNA with 200 U/mL IL-2 protein. Confocal microscopy imaging showing the DAPI-stained nucleus in single images (top) and 3D stacks combining 15 images from top to bottom (middle). 3D stack images show a comparison of nuclear uptake between hIL1β-mRNA and hβactin-mRNA (bottom). d Nuclear signals of hIL1β-mRNA-FITC in CD56⁺CD3^-NK cells treated with siRNA. Control (con) is a nontargeting siRNA (n = 13, 10, and 11 cells for control, ZC3H12D-siRNA and ZC3H12A-siRNA, respectively. Each cell image is composed of 15-stacked 3D images). e IHC analysis of IFN-γ induction 48 h after RNA application with 200 U/mL IL2 to CD56⁺CD3^-NK cells. The positive control was incubated with 20 ng/mL IL12 protein in an IL2 culture medium (IFN-γ signals from n = 16, 21, and 20 ZC3H12D+ cells for no, IL1β-RNA, and IL2 protein, respectively). f IHC analysis of IFN-γ induction 3 and 6 days after stimulation of hIL1β-mRNA and IL2 protein for MDAMB231-TCM-pretreated CD56^dimCD3^-NK cells. The assay system is shown (top). Two donor NK cells were independently used. no, control basal culture medium (bottom left, n = 42, 45, 51, and 51 cells for no, IL1β-RNA, and IL2 protein, and IL1β-RNA plus IL2 protein, respectively. Bottom right, n = 34, 44, 37, and 60 cells for no, IL1β-RNA, and IL2 protein, and IL1β-RNA plus IL2 protein, respectively). Bars, 5 μm. In the graphs, the averages ± SEM, and the results of Student’s t-tests (two-sided) or one-way ANOVA with Bonferroni correction are shown. The P values are shown in the figure. Source data are provided as a Source Data file.