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. 2020 Feb 13;11:860. doi: 10.1038/s41467-020-14678-2

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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. 6 — a Western blotting of B3GALT4-3HA transiently overexpressed in PIGS-UGCG-DKO, PIGS-B4GALT5-DKO, and PIGS-B4GALT5-B4GALT6-TKO cells. b Representative fluorescence images of 3HA-tagged B3GALT4 in PIGS-B3GALT4-DKO, PIGS-UGCG-DKO, and PIGS-B4GALT5-B4GALT6-TKO HEK293 cells. GM130, a marker for cis-Golgi. Scale bar, 10 μm. c Left: PIGS-UGCG-DKO, PIGS-B4GALT5-DKO, and PIGS-B4GALT5-B4GALT6-TKO cells transiently expressing pME-B3GALT4-3HA were stained with T5 mAb. Right: Quantitative data of MFI from four independent experiments (mean ± SD, n = 4). P values are from one-way ANOVA followed by Dunnett’s test for multiple comparisons to PIGS-B3GALT4-DKO cells. d Western blotting of FLAG-6His tagged B3GALT4. Lysates of PIGS-UGCG-DKO, PIGS-B4GALT5-DKO, and PIGS-B4GALT5-B4GALT6-TKO cells stably expressing empty vector (Vec) and B3GALT4 were analyzed. TfR, a loading control. e Left: PIGS-UGCG-DKO, PIGS-B4GALT5-DKO, and PIGS-B4GALT5-B4GALT6-TKO cells stably expressing B3GALT4 were stained with T5 mAb. Right: Quantitative data of MFI from two independent experiments (mean ± SD, n = 2). f Schematic of LacCer enhanced galactose modification on GPI-GalNAc by B3GALT4 in the Golgi. Source data for (c) and (e) are provided as a Source Data file.