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. 2017 Jul 25;7:6474. doi: 10.1038/s41598-017-06622-0

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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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RNA blot-analysis of class-I SlHSP (17.6, 20.0 and 20.1) genes in wild type (Solanum lycopersicum cv. Ailsa Craig) and its ripening mutants. (a) Fruits from wild type tomato var. Ailsa Craig along with near isogenic lines of ripening-inhibitor (rin/rin), non-ripening (nor/nor) and Never-ripe (Nr/Nr) mutant fruits were harvested at mature green [5 days before breaker (−)5BR)], breaker (BR) and red ripe (7 days after breaker, BR + 7) stages as described²⁵. RNA was isolated, separated on gels, blotted and northern blot analysis carried out²⁵, using gene specific probes as previously described⁴⁶. Fruits at mature green stage (−)5BR from the indicated lines were given ethylene treatment and designated as (−)5BR + E. (b) Quantification of northern blot analysis using Image J program (https://imagej.nih.gov/ij/). The band intensity was calibrated with the band intensity of 18 S rRNA internal control used in northern blotting.