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. 2019 Nov 27;9:17692. doi: 10.1038/s41598-019-53544-0

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© The Author(s) 2019

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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Validation of selected genes in Frieseomelitta varia, Melipona quadrifasciata and Scaptotrigona bipunctata. (A) Relative expression of imd was analyzed using the six selected reference genes for qPCR normalization in bees injected with Escherichia coli versus non-injected ones (control, ct). The first and the second most stable reference genes established for at least one of the algorithms (geNorm, NormFinder, BestKeeper, and delta-Ct) are indicated by arrows. Asterisks indicate p < 0.05 (t-test). Bars represent the means and standard errors of three biological replicates. (B) Relative expression (log scale) of abaecin was analyzed using the six selected reference genes for qPCR normalization in bees infected with Escherichia coli (E. coli) versus non-infected ones (control, ct). Asterisks indicate p < 0.05 (t-test). Bars represent the means and standard errors of three biological replicates.