Figure 1.
Image of honeybee pupae in different positions at different developmental stages showing the occurrences of gradual changes in phenotype and organogenesis as time passes.
(A) and (B) represent corresponding whole pupal body and head development.
Figure 2.
Pattern of honeybee pupae head weight at different developmental time points.
Different letters (a, b, c) are significantly different (p<0.05).
Figure 3.
2-DE and CBB G-250 stained gels image showing differentially expressed protein spots profile obtained from different developmental stages of honeybee pupae heads.
Each identified protein spot is specified by number and indicated by arrow, where the spot number with prefix “u” and “d” indicate up or down regulation.
Figure 4.
Functional annotation and distributions of the differentially expressed proteins identified from honeybee pupae head at different developmental time points.
Figure 5.
Functional comparisons of upregulated proteins between the early (13–17 days) and late (19–20 days) stage of honeybee pupae head.
Figure 6.
Global expression intensity map (hierarchical clustering) showing upregulation (red) and downregulation (green) across the ages of the pupae indicated on the top of each column with lists of functional proteins in the right column.
Figure 7.
Functional enrichment analysis of the differentially expressed proteins using the ClueGO software.
Figure 8.
Biological interaction network of the identified differentially expressed proteins from honeybee pupae head at the different developmental stages.
The big ellipse represents identified proteins in this experiment and each entity belongs to the category described in the legend. The “Blue” and “Red” are the proteins upregulated during early to middle development stages (13–17 days) and during late developmental stages (19–20 days), respectively.
Figure 9.
Transcript validation of differentially expressed proteins at different development periods.
mRNA of different development days are measured by quantitative real time PCR. Samples are normalized with reference gene (GAPDH) and with the expression abundant on day 13 as the reference sample. Error bar is standard deviation. Gene symbols indicating different genes refer to Table S2. Different letters (a, b, c) are significantly different (p<0.05).
Figure 10.
Western blot analysis of ald, hsp60, Tcp-1η and idh.
y-axis represents relative expression level normalized by β-actin, x-axis represents different development stages on day 13, 15, 17, 19 and 20, accordingly. Different letters (a, b, c) are significantly different (p<0.05).