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. 2019 Jul 18;9:10471. doi: 10.1038/s41598-019-46952-9

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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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SLC35A1 loss promotes cell death and apoptosis upon VSV infection. (a) Percentage of early apoptotic cells (AnnexinV⁺) and cells that lost membrane integrity due to a late apoptotic/necrotic state (AnnexinV⁺/PI⁺) in HAP1 cell lines expressing sgRNAs against SLC35A1 and Renilla luciferase upon infection with VSV at MOI 2. (b) Immunoblot analysis of Golgi stress signalling pathway activation in the VSV-infected (MOI 2) and Brefeldin A-stimulated (3μg/ml) A549 cells carrying sgRNAs against SLC35A1 and Renilla luciferase. Cropped images are shown for conciseness. Full-length blots are presented in Supplementary Fig. 5. (c,d) VSV-GFP virus replication in SLC35A1 knockout HAP1 cells (c) and HAP1 cells overexpressing SLC35A1 cDNA (d) infected with MOI ranging from 0.001 to 0.01 for 14 hours. (e) Quantification (flow cytometry) of virus titers produced by HAP1wt and two SLC35A1 single cell knockout clones infected with MOI of 2 of VSV-GFP for 8 hours. Statistical significance was assessed using one-way ANOVA with Tukey’s (a,d) and Dunnett’s (c,e) tests. Unless otherwise indicated, adjusted P-values in relation to sgRen control or HAP1wt are shown. Data are represented as mean ± SD of one representative experiment out of at least two independent replicates. **p ≤ 0.01; ***p ≤ 0.001; ns: not significant.