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. 2017 Oct 25;7:13977. doi: 10.1038/s41598-017-13939-3

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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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Representative fields of melanopsin (OPN4) immunostaining in Melan-a (A,B) and B16-F10 (C,D) cells. Cells were kept in DD for 3 days and at the beginning of the 4^th day, cells were divided into 2 groups: (1) Control group kept in constant dark and temperature (37 °C); (2) group in constant dark and exposed to 1 h heat stimulus (39.5 °C). Twenty-four hours later the medium was removed and the cells were fixed with 4% paraformaldehyde. DAPI stained nuclei in blue and OPN4 immunopositivity (1:500 antiserum), revealed with a Cy3-labeled secondary antibody, in orange. Photomicrographies were taken with Axiocam MRm camera (Zeiss) and pseudocolored with Axiovision software (Zeiss). Scale bar 50 μm (200x magnification).