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. 2017 Aug 29;7:9622. doi: 10.1038/s41598-017-08985-w

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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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Immuno-EM micrographs reveal the pattern of penetration of CelA enzymes into pretreated corn stover cell walls and confirm that CelA enzymes occupy the cavities generated in APCS and CFCS. (a,a’) The labeled enzymes (white arrows) in DACS samples appear in cleared zones well into the secondary cell wall and do not appear to be associated with surface lignin globules. (b,b’,c,c’) CelA enzyme labeled in APCS and CFCS cell walls were most often found within cavities that connect to the cell wall surface and penetrate into the secondary cell wall. (d) Quantitation of the immune gold α-CelA CBM3 labeling density shows 3–4 times the labeling density in the digested APCS or CFCS compared to DACS.