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. 2020 May 18;10:8137. doi: 10.1038/s41598-020-65035-8

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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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Intracellular delivery of scFv by CPP in SH-SY5Y and HEK293T cells. SH-SY5Y (a,b) or HEK293T (c,d,e) cells incubated with scFv-pF and -pC for 120 min in Opti-MEM were fixed and stained using anti-His antibody and Phalloidin-594 and imaged using confocal microscope. Serial optical sections in the Z-axis of the cell, collected at 1 μm intervals with 63x oil immersion objective lense (NA 1.4) were projected and observed in a total thickness of 10 μm by using LSM 780 (version 3.2) software. scFv-pF (a or c) is not showing any specific staining indicating no intracellular delivery due to the lack of any cell penetrating peptide. scFv-pC (b,d) is showing good specific staining around the cells and inside cells indicating its binding to the cell membrane and possibly intracellular delivery due to the presence of CPP-peptide. (e) Merged confocal image with the orthogonal projection of scFv-pF (left) and scFv-pC (right) localization along with XZ and YZ views. Scale bars = 10 μm.