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Fig. 4 | BMC Ecology and Evolution

Fig. 4

From: Evolution and genomic organization of the insect sHSP gene cluster and coordinate regulation in phenotypic plasticity

Fig. 4

CTCF defines H3K27me3 enrichment at the boundaries of the sHSP-C. A The genomic structure at the sHSP-C in honeybees. Regulatory motifs CTCF (blue dotted line), Br-C (yellow circles) and HSE (green circles). B Average H3K27me3 enrichment across the sHSP-C. Cyan is queen H3K27me3 enrichment, black is from queen-less workers, and magenta is from queen-right workers. The sHSP-C bound by CTCF (including Gmap and LOC100576174) is significantly depleted of H3K27me3 in queen-right worker, queen-less worker and queen ovaries (Kruskall Wallis with a Dunn post-hoc test) in comparison to the 5ʹ and 3ʹ flanks. Numbers denote statistical significance (p < 0.0001). 1, 3 and 5 are comparisons between the 5ʹ flank and CTCF bound cluster for queen-right, queen-less worker and queens respectively. 2, 4 and 6 are comparisons between the 3ʹ flank and CTCF bound cluster for queen-right worker, queen-less worker and queen respectively. C Boxplot illustrating H3K27me3 enrichment across the sHSP-C and the 3ʹ and 5ʹ flanks that are demarcated by CTCF. Boxplot whiskers indicate minimum and maximum, the box is defined by 25th percentile, median, and 75th percentile. Outliers, data points outside 1.5 times the interquartile range above the upper quartile and below the lower quartile, are shown as individual data points. D RT-qPCR of genes honeybee sHSP-C as well as immediately adjacent genes Gmap and LOC100576174. Target gene expression is measured relative to mRPL44 and Rpn2, which are stably expressed in honeybee ovaries [54]. Gene expression was measured in five biological replicates each consisting of ovaries from multiple individuals: queen (n = 3), queen-right worker (n = 20), and queen-less workers (n = 10). Differences in gene expression were assessed using a general linear model ANOVA with a Tukey post hoc test and 95% confidence interval. Samples that do not share letters are statistically significantly different with a p value < 0.05. All of the sHSP-C genes are significantly different in expression between queen-right worker and queen-less worker ovaries (with the exception of LOC724274) whereas the genes Gmap and LOC100576174 which are immediately adjacent to the sHSP-C are not significantly different in expression between queen-right worker and queen-less worker ovaries. This data validates what we see in the ovary activation RNA-seq dataset comparing sHSP-C gene expression in queen-right worker and queen-less worker ovaries

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