Spearman correlation network for samples treated with LPS at time point 2h.

<p>(A) The complete network showing interaction between genes at 0 hours after LPS treatment. Highly connected hubs includes <i>TLR4</i>, <i>IFNB</i>, <i>MyD88</i>, <i>IL8</i>, and <i>HSPA14</i>. (B) Cluster 1 from MCODE network clustering algorithm showing the interaction between <i>IL8</i>, <i>IL1B</i>, <i>CCL5</i>, <i>CCL4</i>, <i>iNOS</i>, <i>JUN</i>, and HSPA14 after ignoring the hairball to the right of <i>HSPA14</i>. The cluster shows interaction between immune response genes from LPS stimulation.</p

Spearman correlation network for samples treated with at time point 2h.

<p>(A) The complete network showing interaction between genes at 0 hours after heat stress treatment. Highly connected hubs includes <i>iNOS</i>, <i>IL8</i>, <i>CD40</i>, and <i>HSPA14</i>. (B) Cluster 2 from MCODE network clustering algorithm showing the interaction between <i>iNOS</i>, <i>IFNB</i>, <i>CASP9</i>, <i>CASP3</i>, <i>HSPA14</i>, <i>IFNG</i>, and <i>MAPK9</i>. The cluster shows interaction between multiple mechanisms during response to heat stress.</p

Temporal changes in the relative expression of the genes showing the highest up- or down-regulation upon heat stress and LPS treatment.

<p>Fold induction in the relative gene expression was calculated using ddCt method (n = 9). The genes were organized by their function in: heat stress response (<i>HSP25</i>, <i>HSPA2</i>, <i>HSPH1</i>, <i>DNAJA4</i> and <i>DNAJB6</i>), stress response (<i>BAG3</i> and <i>UBB</i>) and immune response (<i>CCL4</i>, <i>CCL5</i>, <i>IL1B</i>, <i>IL8</i> and <i>iNOS</i>). X-axis presents a time point (2h, 4h, 6h and 10h). Y-axis shows fold induction of the gene expression (logarithmic scale). Untreated (control) samples calibrated at the value of fold induction = 1.</p

Function of the target genes used in the gene expression study.

<p>Function of the target genes used in the gene expression study.</p

Spearman correlation network for samples treated with at time point 2h.

<p>(A) The complete network showing interaction between genes at 0 hours after heat stress treatment. Highly connected hubs includes <i>iNOS</i>, <i>IL8</i>, <i>CD40</i>, and <i>HSPA14</i>. (B) Cluster 2 from MCODE network clustering algorithm showing the interaction between <i>iNOS</i>, <i>IFNB</i>, <i>CASP9</i>, <i>CASP3</i>, <i>HSPA14</i>, <i>IFNG</i>, and <i>MAPK9</i>. The cluster shows interaction between multiple mechanisms during response to heat stress.</p

Spearman correlation network for samples treated with LPS and heat stress samples at time point 2h.

<p>(A) The complete network showing interaction between genes at 0 hours after LPS under heat stress treatment. Highly connected hubs includes <i>CASP9</i>, <i>IL18</i>, <i>IFNB</i>, <i>IL8</i>, and <i>iNOS</i>. There appears to be a small cluster of genes related to apoptotic pathway formed near the top of the network. (B) Cluster 2 from MCODE network clustering algorithm showing the interaction between <i>IL8</i>, <i>IL18</i>, <i>TLR4</i>, <i>TRAF6</i>, <i>CD40</i>, <i>IFNB</i>, <i>IFNG</i>, and <i>CASP9</i>. The cluster shows interaction between multiple mechanisms during response to LPS stimulation under heat stress. Interesting to note once again the additional interaction with the <i>CASP9</i>/<i>CASP3</i> apoptotic pathway.</p

Log₂ Fold Change Heat Map.

<p>A heat map for the 3 different treatments across the 4 time points. Log₂ fold change calculate based on delta Ct value compared to the control samples and green implies increased expression while red implies decreased expression. Genes on the right are clustered using a hierarchical clustering method and 8 clusters were found. Cluster 1 = <i>BAG3</i>, <i>HSPA2</i>, <i>HSPH1</i>, <i>DNAJA4</i>, <i>HSP25</i>; Cluster 2 = <i>iNOS</i>, <i>IL1B</i>, <i>CCL5</i>, <i>CCl4</i>, <i>IL8</i>; Cluster 3 = <i>IL12B</i>, <i>IFNG</i>, <i>UBB</i>; Cluster 4 = <i>TGFB2</i>, <i>DNAJB6</i>, <i>JUN</i>, <i>HSP90</i>; Cluster 5 = <i>TLR4</i>, <i>NLRC5</i>, <i>HSF2</i>, <i>HSF5</i>; Cluster 6 = <i>CASP8</i>, <i>IL18</i>, <i>RB1CC1</i>, <i>TRAF6</i>, <i>SMAD6</i>, <i>MyD88</i>, <i>CASP7</i>, <i>CASP1</i>, <i>IRAK4</i>, <i>TP53</i>; Cluster 7 = <i>CASP9</i>, <i>MAP48IP3</i>, <i>CASP3</i>; Cluster 8 = <i>CIRBP</i>, <i>MAPK9</i>, <i>IFNB</i>, <i>TGFB3</i>, <i>HSPA14</i>, <i>LITAF</i>, CD40. Each gene is color coded based on their major functional category: heat stress response (red), immune response (green), signaling (yellow), and stress response (orange).</p