Quantitative Real Time PCR (qRT-PCR) validation of DGE result.

a<p>means significant difference existed between the DA treated and control samples.</p>b<p>means no significant difference.</p><p>DGE data were generated from two biological repeats. qPCR data were generated by the standard deviation from three biological repeats.</p

Gene Expression Profile of <i>Bombyx mori</i> Hemocyte under the Stress of Destruxin A

<div><p>Destruxin A (DA) is a cyclo-peptidic mycotoxin from the entomopathogenic fungus <i>Metarhizium anisopliae</i>. To uncover potential genes associated with its molecular mechanisms, a digital gene expression (DGE) profiling analysis was used to compare differentially expressed genes in the hemocytes of silkworm larvae treated with DA. Ten DGE libraries were constructed, sequenced, and assembled, and the unigenes with least 2.0-fold difference were further analyzed. The numbers of up-regulated genes were 10, 20, 18, 74 and 8, as well as the numbers of down-regulated genes were 0, 1, 8, 13 and 3 at 1, 4, 8, 12 and 24 h post treatment, respectively. Totally, the expression of 132 genes were significantly changed, among them, 1, 3 and 12 genes were continually up-regulated at 4, 3 and 2 different time points, respectively, while 1 gene was either up or down-regulated continually at 2 different time points. Furthermore, 68 genes were assigned to one or multiple gene ontology (GO) terms and 89 genes were assigned to specific Kyoto Encyclopedia of Genes and Genomes (KEGG) Orthology. In-depth analysis identified that these genes putatively involved in insecticide resistance, cell apoptosis, and innate immune defense. Finally, twenty differentially expressed genes were randomly chosen and validated by quantitative real-time PCR (qRT-PCR). Our studies provide insights into the toxic effect of this microbial insecticide on silkworm's hemocytes, and are helpful to better understanding of the molecular mechanisms of DA as a biological insecticide.</p></div

Quality assessment of reads in each library.

<p>Quality assessment of reads in each library.</p

Summarizing of Gene Ontology functional classification of ten DGEs by comparing DA-treated and un-treated samples in differential time intervals, which described gene products in terms of their associated biological processes, cellular components and molecular functions.

<p>Summarizing of Gene Ontology functional classification of ten DGEs by comparing DA-treated and un-treated samples in differential time intervals, which described gene products in terms of their associated biological processes, cellular components and molecular functions.</p

Screening of more than 2-fold differentially expressed genes in <i>B.mori</i> hemocytes after the treatment of DA in differential time intervals.

<p>Screening of more than 2-fold differentially expressed genes in <i>B.mori</i> hemocytes after the treatment of DA in differential time intervals.</p

NCBI SRA accession numbers for the treatments at each time point.

<p>NCBI SRA accession numbers for the treatments at each time point.</p

Statistics of mapping to reference transcriptome for each library.

<p>Statistics of mapping to reference transcriptome for each library.</p

The MALDI-TOF/TOF-MS/MS analysis of protein spot 22.

<p>Note: The MALDI-TOF-MS peptide mass fingerprint spectrum of trypsin-digested protein (a) and its MS/MS peptide mass fingerprint spectrum of ionic peak 2565.23 (b).</p

qRT-PCR validation of DGE result.

<p>Note: The left y-axis indicates the relative expression level by qRT-PCR, and the right y-axis indicates the log₂Ratio of 4H/CK by DGE.</p

Western blot analysis of expression of <i>Px</i>Serpin 2.

<p>Note: Those were visualized by DAB. Actin was used as an internal control.</p