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. 2017 Nov 21;7:15901. doi: 10.1038/s41598-017-15567-3

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© The Author(s) 2017

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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VacA-dependent and VacA-independent increase of POLG and TFAM early upon H. pylori infection. AGS cells co-cultured with H. pylori 26695 or 26695∆vacA. (A) POLG and (D) TFAM immunofluorescence (red, left panels); merge with TOM22 immunostaining (green) and nuclei (Hoechst, blue) (right panels). Scale bar: 10µm. (B) POLG and (E) TFAM quantification of immunofluorescence intensity (n = 30 cells/condition (and n = 50 non-infected cells) from three independent experiments mean ± SD, Welch’s test, **p < 0.01; ***p < 0.001; ****p < 0.0001. WB of (C) POLG and (F) TFAM (human 29 kDa and 25 kDa isoforms, position indicated with a star). Samples were derived from the same experiment and the gels and blots were processed in parallel (one gel for 2 h and 6 h, one gel for 24 h and 48 h). Each blot was then sliced in horizontal sections to allow multiple immunolabelings with the same membrane. Sections were alternatively labeled with anti-POLG and anti-TFAM (other labelings performed with the remaining portions of the membrane are not shown). The corresponding uncropped gels/blots are shown in Supplementary Figure S7C. SYPRO Ruby blot staining used as loading control (total protein staining is used as alternative to overcome single housekeeping protein variations) is shown in Supplementary Figure S7C (entire gel staining). Data from 3 independent experiments.