Fig. 4

Integrated transcriptomic analysis of the onset, maintenance and termination of diapause. A Schematic diagram shows the RNA-Seq workflow time course. B PCA displays gene expression profiles across all sequenced samples. C Scatterplot for comparing the fold change of gene expression during the reactivation process. Each dot represents a DEG identified by comparing pre-diapause embryos (F0) with diapause embryos (S2d). The x-axis shows the log2fold change in expression in pre-diapause embryos (F0) compared with diapause embryos (S2d). The y-axis shows the log2fold change of expression in reactivating embryos (In0.5h–In12h) compared with diapause embryos (S2d). Linear regression analysis was performed for each panel. The Pearson R, p value and slope of the regression line are indicated. D Boxplots show the expression profile of genes that are annotated in cell proliferation and the cell cycle. The y-axis represents the log10 fold change. The horizontal blue line is the zero line of log10 (fold change). Black lines in the boxes indicate the median of log10 (fold change), and the + symbols in the boxes indicate the mean of log10 (fold change). A gene set comprising 150 random selected genes served as a control. E RNA-Seq data show dynamic changes in expression of genes at different time points. The dashed line indicates the expression level in F0 embryos, N = 3. F qPCR of the changes in gene expression after embryonic stem cells were induced by cold stress (28°C) for 3 hours (C3h) or 6 hours (C6h), N = 5. *p < 0.05; **p < 0.01; ***p < 0.001. N indicates biological replicates