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. 2017 Aug 17;7:8531. doi: 10.1038/s41598-017-09067-7

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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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Screening for a protein domain/subdomain that controls protein folding/cellular quality system. (a) Illustrates screening for a sequence to improve the stability of a protein of interest by incorporation of protein domains or subdomains from various chaperone or co-chaperone proteins. (b) Shows X-ray film images of chemiluminescent signals of a western blot (immunoblot) analysis of cell lysates, indicating relative levels of the expression of V5-tagged IL13Rα2 wild-type (IL13Rα2-V5) protein or fusion protein counterparts (IL13Rα2-J-domain fragment-V5) in transfected cells. HEK293 cells were transfected with an empty vector (lane 1); the plasmid for IL13Rα2 alone (lane 2); fusion protein of IL13Rα2 with a J-domain sequence derived from DNAJB1 (J^DNAJB1; lane 3); CSP (J^DNAJC5; lane 4); and SV40 large T antigen (J^SV40; lane 5). All transfectants were co-transfected with a reporter plasmid expressing green fluorescent protein (GFP) to demonstrate the successful transfection and operability of the transfectants (lower panel). (c) HEK293 cells were transfected with an empty vector (lane 1); a plasmid for a V5-tagged truncated form (“TF”) of IL13Rα2 receptor protein (IL13Rα2TF-V5; lane 2); the extracellular portion of the IL13Rα2 receptor protein containing the ligand binding domain; and a fusion protein of IL13Rα2TF with a J-domain fragment sequence from Erdj3 (IL13Rα2TF -J^erdj3-V5; lane 3). Two days later, the cell culture medium (right panel) and cell lysate (left panel) were harvested, and western blot analysis was conducted with an anti-V5 antibody. A reporter plasmid (GFP) was co-transfected to monitor transfection efficiency (lower panel). (d) J-domain sequence was inserted between signal sequence and a V5-tagged IL13Rα2TF and expressed in HEK293 cells. After the expression, the culture medium (top panel) and cell lysate (middle panel) were harvested to analyze the expression of the protein. Lane1: mock transfectant, lane2: IL13Rα2TF-V5, and lane 3 shows the N-terminal fusion protein with the J-domain sequence (J^erdj3-IL13Rα2TF-V5). GFP was co-transfected for transfection efficiency (lower panel).