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. 2018 Jan 23;8:1401. doi: 10.1038/s41598-018-19831-y

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© The Author(s) 2018

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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Western blots of pGH protein from strains contained P_AOX1 variants. For detection of pGH, 50 μg intracellular proteins were used for SDS-PAGE while Rabbit anti-pGH polyclonal antibody was used for Western blot. The pGH expression level in poly(dA:dT) tracts deletion variants were shown in Lane 3–9 (a), Lane 1 (b) and Lane 1–9 (c); The pGH expression level in addition variants were shown in Lane 2–9 (b) and Lane 1–9 (d). CK represented negative control samples from pPICZA transformant; WT represented the pGH expression level from strain harboring wild-type AOX1 promoter. The relative expression levels of pGH in all strains were quantified with ImageJ software. WT was chosen as the standard sample to perform relative comparison. The integrated density of each band represented the abundance of protein. The results of three independent cultivations were indicated as mean ± SD. The relative expression level of pGH in deletion variants were shown in e and addition variants in f.