One particular.0255125 September 7,11 /PLOS ONEPentosan polysulfate sodium prevents functional decline in chikungunya infected miceFig 6. qPCR NanoStringTM panel and bioinformatic evaluation of muscle tissue. Gene NKG2C/CD159c Proteins Formulation expression analysis of RNA isolated from muscle tissue at peak disease was performed using the commercially readily available NanoStringTM nCounter1 mouse Myeloid Innate Immunity gene expression panel. Data shows (A) volcano plots, (B) heat maps and (C) box plots (Log2) of key genes which are modulated in quadriceps during PPS remedy. Gene expression information was determined applying Limma package and P values have been adjusted for multiple testing by Benjamini and Hochberg approach to manage the false discovery price. Data compares tissues from CHIKV-infected PPS-treated mice to CHIKV-infected untreated mice (n = three mice/group). Prime genes selected had a FC 1.three or FC -1.three in addition to a P value 0.02. https://doi.org/10.1371/journal.pone.0255125.gPLOS A single https://doi.org/10.1371/journal.pone.0255125 September 7,12 /PLOS ONEPentosan polysulfate sodium prevents functional decline in chikungunya infected miceFig 7. Protein-protein interaction network. PPI network was generated making use of the top hits of each joint and muscle tissues combined. The reduce off criteria included at the least a 1.three FC and a P value 0.02. Circles represent genes and lines represent the interaction of proteins amongst genes. Line colour represents evidence of interaction among the proteins. Prime hits were selected according to lowest FDR. Benefits for both KEGG and REACTOME pathways are shown. Major three functional groups as identified with NanoStringTM annotations are also shown. https://doi.org/10.1371/journal.pone.0255125.ginteraction, which once again suggest essential roles inside the effects observed with PPS treatment through CHIKV infection. General, the PPI network provides fundamental insights into the important targets of PPS therapy of CHIKV-induced inflammatory illness. Results of key KEGG and REACTOME pathways were also generated making use of STRING evaluation. The top rated five outcomes, based on FDR and identified via the KEGG pathway had been cytokinecytokine receptor interactions, pathways in cancer, phosphatidylinositol 3-kinase PI3-AKT signalling, epidermal development aspect receptor (EGFR), tyrosine kinase inhibitor resistance and hypoxia inducible factor (HIF) signalling pathway. For REACTOME, the top 5 pathways incorporated Flk-1/CD309 Proteins Source signal transduction pathways, signalling by tyrosine-protein kinase 6 (PTK6), mitogen-activated protein kinase 1 and three (MAPK1/MAPK3) signalling, signalling by receptor tyrosine kinases and peptide-ligand-binding receptors (FDR 0.0004; Fig 7). Major DEGs were also sorted making use of functional group evaluation. NanoStringTM delivers functional annotations to classify DEGs of a specific panel into biological course of action categories. The top 3 functional groups integrated: development element signalling (Il-15ra, Il-1, Cdkn1a, Il-6ra, Egf, tyrosine-protein kinase Met (Met), nuclear aspect of activated T cell 2 (Nfatc2), tyrosineprotein phosphatase non-receptor form 14 (Ptpn14), Stat1, Vegfa, histone deacetylase 5 (Hdac5), Ras-specific guanine nucleotide-releasing factor two (Rasgrf2) and Fgf2), lymphocyte activation (Clec7a), adenosine A3 receptor (Adora3), Cdkn1a, Fcer1a, formyl peptide receptorrelated sequence 3 (Fpr-rs3), apoptosis-associated speck-like protein containing a CARD (Pycard; ASC), Egf, Fgf2, Nfatc2, Semaphorins 6D (Semad6), Il-1 and Rasgrf2) and pathogenPLOS 1 https://doi.org/10.1371/journal.pone.0255125 September 7,13 /.