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. 2019 Mar 11;10(3):236. doi: 10.1038/s41419-019-1487-3

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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 — a Western blot analysis of protein extracts from the indicated transiently transfected HCT116 cells using antibodies against NAA40, H4R3me1, H4R3me2a, H4R3me2s, total histone H4, and β-actin as loading control. The densitometry numbers below each blot define the normalized levels of H4R3me1, H4R3me2a, and H4R3me2s against H4 and of NAA40 against β-actin relative to SCR cells. b Western blot analysis of protein extracts from the indicated siRNA-treated cells using antibodies toward NAA40, PRMT5, PRMT7, and β-actin as loading control. The values below each blot were calculated by densitometry analysis of NAA40, PRMT5, and PRMT7 bands relative to SCR control after normalization with β-actin. c qRT-PCR analysis of mRNA levels of NAA40, PRMT5, and PRMT7 normalized to β-actin in the indicated HCT116 cells. Error bars represent the mean ± s.d of three biological replicates. Unpaired two-tailed Student’s t-test was used (ns = no significance, **p < 0.01, ***p < 0.001). d ChIP assay monitoring the enrichment of N-acH4, H3K4me3, and H3K27me3 on the PRMT5 gene promoter. The enrichment from each antibody was normalized to total histone H3. Results are shown as mean ± s.d. from two independent experiments. Unpaired two-tailed Student’s t-test was used (**p < 0.01, ***p < 0.001). e qRT-PCR analysis of mRNA levels in HCT116 cells for the indicated genes. The results were normalized to β-actin mRNA and represent the average from three independent experiments. Unpaired two-tailed Student’s t-test was used (ns = no significance, **p < 0.01, ***p < 0.001)