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. 2022 May 21;8:48. doi: 10.1038/s41421-022-00395-1

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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. 4 — a Quantitative RT-PCR analysis of precursor or mature TM9SF1 mRNA expression in AGS cells with control or PCIF1 depletion. Data are presented as log₂ value of PCIF1 relative to GAPDH. b Analysis of remaining TM9SF1 mRNA at the indicated times in control and PCIF1-depleted AGS cells after actinomycin D treatment. c–f The expression of m⁷G-Am or m⁷G-m⁶Am EGFP mRNA in Hela cells. EGFP mRNA that starts with either m⁷G-Am or m⁷G-m⁶Am modification was generated by in vitro transcription (c). Cells were transfected with m⁷G-Am or m⁷G-m⁶Am EGFP mRNA and subjected to qRT-PCR (d) and fluorescence microscopy (e). Representative images of EGFP in cells are shown. Fluorescence intensity of EGFP-positive Hela cells was quantified (f). DNA was visualized by DAPI. g, h Polysome profiling analysis of TM9SF1 mRNA. The distribution of TM9SF1 mRNA in each fraction of control and PCIF1-depleted BGC-823 cells was determined by qRT-PCR. i Analysis of TM9SF1 mRNA distribution in the fractions of non-polysome and polysome from BGC-823 cells with the indicated siRNA and plasmids. j Western blot analysis of TM9SF1 in AGS cells with the indicated shRNA and plasmids. Data are shown as means ± SD. N.S. not significant; *P < 0.05, **P < 0.01, Student’s t-test.