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. 2021 Dec 8;12:7142. doi: 10.1038/s41467-021-27525-9

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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. 5 — a RNA-seq was performed in PLC-8024-CTR and PLC-8024-GDF1 cells. Gene set enrichment analysis (GSEA) was performed in PLC-8024 cells transfected with or without GDF1 to examine the enrichment in gene sets of cancer-testis antigens. NES, normalised enrichment score. b The relative expression of representative CTAs was detected by qPCR in PLC-8024-CTR and PLC-8024-GDF1 cells (*P < 0.05, **P < 0.01, ***P < 0.001, NS, not significant, two-tailed independent Student’s t test, data are presented as mean values ± SD, n = 3 independent experiments). c The relative expression of representative CTAs was detected by qPCR in PLC-8024 cells cocultured with PLC-8024-CTR and PLC-8024-GDF1 cells (**P < 0.01, ***P < 0.001, NS, not significant, two-tailed independent Student’s t-test, data are presented as mean values ± SD, n = 3 independent experiments). d The relative expression of representative CTAs was detected by qPCR in PLC-8024 cells treated with rGDF1 at 50 ng/mL or vehicle control for 15 days (*P < 0.05, ***P < 0.001, NS not significant, two-tailed independent Student’s t test, data are presented as mean values ± SD, n = 3 independent experiments). e IHC staining of GAGE12E in subcutaneous and intrahepatic tumours formed by PLC-8024-CTR and PLC-8024-GDF1 cells. Scale bars represent 50 µm, n = 3 independent experiments. f The relative expression of representative LSD1 was detected by qPCR in PLC-8024 and Huh7 cells transfected with or without GDF1 (two-tailed independent Student’s t-test, data are presented as mean values ± SD, n = 3 independent experiments). g PLC-8024 and Huh7 cells were treated with the LSD1 inhibitor GSK-LSD1 at 5 μM for 72 h. The relative expression of representative CTAs was detected by qPCR (*P < 0.05, **P < 0.01, ***P < 0.001, NS, not significant, two-tailed independent Student’s t test, data are presented as mean values ± SEM, n = 3 independent experiments). h PLC-8024-CTR and PLC-8024-GDF1 cells were treated with LSD1 inhibitor at 5 μM or 10 μM for 72 h. The expression of GAGE12E and LSD1 at the protein level was examined by western blot. iLSD1, GSK-LSD1. + , 5 μM. ++,10 μM (n = 3 independent experiments). i PLC-8024 cells were treated with rGDF1 at 50 ng/mL or TGF-β1 at 10 ng/mL or vehicle control for 30 min. The enrichment of SMAD2/3 on LSD1 promoter under stimulation with either recombinant GDF1 or TGF-β1 was conducted by ChIP-PCR assay (two-tailed independent Student’s t-test, data are presented as mean values ± SEM, n = 3 independent experiments). j Promoter activities of LSD1 in PLC-8024 cells treated with rGDF1 at 50 ng/mL or TGF-β1 at 10 ng/mL or vehicle control for indicated time points were detected by dual-luciferase assay (two-tailed independent Student’s t-test, data are presented as mean values ± SEM, n = 3 independent experiments). iLSD1, GSK-LSD1. rGDF1, recombinant human GDF1 protein. rTGFB1, recombinant human TGF-β1 protein. k Ectopic expression of SMAD3 was induced in GDF1-knockout cell lines (n = 3 independent experiments). The expression of GAGE12E and LSD1 at the protein level was examined by western blot. Source data are provided as a Source Data file.