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. 2020 Mar 10;6:12. doi: 10.1038/s41421-020-0144-4

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© The Author(s) 2020

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. 2 — a, b qRT-PCR and immunoblot analysis of YTHDF3 mRNA (a) and protein (b) in trophoblast treated with hypoxia as indicated time. The gene expression was normalized to that of the β-actin internal control in each sample. c, d qRT-PCR and immunoblot analysis of YTHDF3 in trophoblast HTR8/SVneo transfected with lentiviral shControl (NC), YTHDF3 shRNA #1 or #2 as indicated for 72 h. e, f Left: Migration (e) and invasion (f) analysis of HTR8/SVneo cells stably expressing control shRNA (shControl) vs. shRNA targeting YTHDF3 (shYTHDF3) by Transwell assay. Representative images are shown. Right: The number of migration and invasion cells was counted by the ImageJ software. g, k Cell proliferation measured by CCK8 assay of HTR8/SVneo cells stably expressing shYTHDF3 (g) and YTHDF3 (k) as indicated. The value of OD₄₅₀ was normalized to the value about 24 h after cell seeding. h Immunoblot analysis of HTR8/SVneo cells stably expressing YTHDF3 as indicated. i, j Left: Migration (i) and invasion (j) analysis of HTR8/SVneo cells transfected with YTHDF3 plasmid by Transwell assay. Representative images are shown. Right: The number of migration and invasion cells was counted by the ImageJ software. l qRT-PCR analysis of MMP9 and MMP2 mRNA in HTR8/SVneo stably expressing shControl and shYTHDF3 as indicated. Scale bars, 100 μm; *p < 0.05, **p < 0.01 (Student’s t test). Data are representative of three independent experiments (mean and s.d. of technical triplicates (a, c, g, k, l)).