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Figure 1

A. The PQ mice displayed a collapse of pulmonary architecture, severe infiltration of mononuclear cells into intra-alveolar and interstitial spaces, and edematous interstitial space. B. The histopathological results reflected that lung injuriesy was were triggered at 6 hr and 24 hr after PQ, which corresponded to the histopathological specimen (p<0.01 and p<0.05 respectively). 

 

Figure 2 

By In utilizing a microarray analysis, the heat map showed that a total of 2287 genes were identified that showed statistically significant differential expression in PQ mice at 6h and 24h when compared to controls. 

 

Figure 3 

A. By using STEM software identified, 16 profiles containing a cluster of multiple genes which have similar expression patterns after PQ were identified (Figure 3). Five of these clusters are comprised of genes that were repressed gradually express different expression levels through the time points, for example profiles 9, 4, 7 13 and 15; while genes in profiles 4 and 1 had opposite effects. B. The profiles of 4 and 7 shared the a very similar descending expression at 6hr and 24 hr after PQ. 

 

Figure 4 

By GO-analysis, in the patterns of 4 and 7 which genes have descending expression duringover the time course of 6 hr and 24 hr, the highest enrichments of GO are all associated with mitochondron.

 

Figure 5

A. 76 genes wereas selected as the candidate genes from the total differential genes as function of mitochondron by constructing a gene co-expression network with k-core algorithm. These 76 genes all belongs into STC patterns 4 and 7.

B. In the co-expression network of 76 genes of pattern 4 and 7, PSTK gene localiseslocalizes at the center of 11 k-core subnetwork. It directly regulates 30 neighboring genes that interact according to their degrees. These interactions depend in large part on PSTK because the clustering coefficients of these genes are above 0.99, which are higher than for other genes. 

C and D. A heat map showed these 31 differential genes and verified that the expression of PSTK mRNA is gradually decreased.

 

Figure 6 

A. Real-time RT-PCR revealed that the mRNA level of PSTK showed a statistically significant decrease (p < 0.05) in lungs at 6hrs and 24 hrs, when compared with that those from control mice. But there was no statistical significance (p > 0.05) between 24 hrs and 6 hrs groups.

B. Western Blot showed that tThe PSTK protein level was less expressed in both 6hr and 24 hr groups as compared with that in normal groups. But tThere was, however, no difference between 6 hr and 24 hr groups.

 

Figure 7 

A. ImmunofluorenceImmunofluorescence staining showed that PSTK in lungs has a less lower number in lungs from 6hr and 24 hr groups as compared with the normal groups. A. Under the HPF magnitude microscopy, the number of PSTK positive staining (the green dots) from normal control groups has nearly two times more than that from both 6 hr and 24 hr PQ groups. 

B. Quantification of PSTK immunofluorenceimmunofluorescence staining in normal control and PQ groups.