Journal: PNAS Nexus

Article Title: Vascular endothelial cell-specific disruption of the profilin1 gene leads to severe multiorgan pathology and inflammation causing mortality

doi: 10.1093/pnasnexus/pgad305

Figure Lengend Snippet: Impact of Pfn1 depletion on EC transcriptome. A) Heat plot showing differentially expressed genes between Ad-GFP– and Ad-Cre–infected EC (genes color coded by blue and yellow denote transcriptionally downregulated and upregulated genes in Ad-Cre relative to Ad-GFP–infected cells; data summarized from three biological replicates). B) Heat plot showing transcriptionally altered cytokines/chemokines as a result of Pfn1 depletion in EC (numbers alongside indicate the average fold change in Ad-Cre relative Ad-GFP groups; asterisks denote robustly altered cytokines/chemokines that are in agreement with serum and/or CM Luminex data). C) A volcano plot ( P -value vs Z score) displaying IPA-predicted activated and inhibited transcriptional regulators in Ad-Cre– vs Ad-GFP–infected cells. Top IPA-predicted activated transcription factors (based on the absolute Z score and negative log P -value) are outlined by a green oval. D) Representative immunoblots showing the relative levels of the total as well as phosphorylated forms of STAT1 and IRF7 between Ad-GFP– and Ad-Cre–infected cells (GAPDH blot serves as the loading control; Pfn1 blot confirms Ad-Cre–induced suppression of Pfn1 expression).

Article Snippet: TCL was run on an SDS–PAGE and immunoblotted using antibodies specific for Pfn1 (Abcam, ab124904; 1:3,000), IRF7 (Cell Signaling Technology, Danvers MA, 72073; 1:1,000), p-IRF7 (Cell Signaling Technology, 24129; 1:1,000), STAT1 (Cell Signaling Technology, 9172; 1:1000), p-STAT1 (Cell Signaling Technology, 9167; 1:1,000), and GAPDH (Sigma-Aldrich, G9545; 1:2,000).

Techniques: Infection, Luminex, Western Blot, Control, Expressing

Journal: PNAS Nexus

Article Title: Vascular endothelial cell-specific disruption of the profilin1 gene leads to severe multiorgan pathology and inflammation causing mortality

doi: 10.1093/pnasnexus/pgad305

Figure Lengend Snippet: Schematic representation of how endothelial Pfn1 loss could potentially lead to a proinflammatory condition. We propose that loss of Pfn1 in EC directly stimulates select proinflammatory cytokine/chemokine expression/release through upregulating the intrinsic IRF7/STAT1 signaling in addition to potentiating proinflammatory gene expression in monocytes/macrophages in a paracrine signaling manner.

Article Snippet: TCL was run on an SDS–PAGE and immunoblotted using antibodies specific for Pfn1 (Abcam, ab124904; 1:3,000), IRF7 (Cell Signaling Technology, Danvers MA, 72073; 1:1,000), p-IRF7 (Cell Signaling Technology, 24129; 1:1,000), STAT1 (Cell Signaling Technology, 9172; 1:1000), p-STAT1 (Cell Signaling Technology, 9167; 1:1,000), and GAPDH (Sigma-Aldrich, G9545; 1:2,000).

Techniques: Expressing, Gene Expression

Journal: PNAS Nexus

Article Title: Vascular endothelial cell-specific disruption of the profilin1 gene leads to severe multiorgan pathology and inflammation causing mortality

doi: 10.1093/pnasnexus/pgad305

Figure Lengend Snippet: Impact of Pfn1 depletion on EC transcriptome. A) Heat plot showing differentially expressed genes between Ad-GFP– and Ad-Cre–infected EC (genes color coded by blue and yellow denote transcriptionally downregulated and upregulated genes in Ad-Cre relative to Ad-GFP–infected cells; data summarized from three biological replicates). B) Heat plot showing transcriptionally altered cytokines/chemokines as a result of Pfn1 depletion in EC (numbers alongside indicate the average fold change in Ad-Cre relative Ad-GFP groups; asterisks denote robustly altered cytokines/chemokines that are in agreement with serum and/or CM Luminex data). C) A volcano plot ( P -value vs Z score) displaying IPA-predicted activated and inhibited transcriptional regulators in Ad-Cre– vs Ad-GFP–infected cells. Top IPA-predicted activated transcription factors (based on the absolute Z score and negative log P -value) are outlined by a green oval. D) Representative immunoblots showing the relative levels of the total as well as phosphorylated forms of STAT1 and IRF7 between Ad-GFP– and Ad-Cre–infected cells (GAPDH blot serves as the loading control; Pfn1 blot confirms Ad-Cre–induced suppression of Pfn1 expression).

Article Snippet: TCL was run on an SDS–PAGE and immunoblotted using antibodies specific for Pfn1 (Abcam, ab124904; 1:3,000), IRF7 (Cell Signaling Technology, Danvers MA, 72073; 1:1,000), p-IRF7 (Cell Signaling Technology, 24129; 1:1,000), STAT1 (Cell Signaling Technology, 9172; 1:1000), p-STAT1 (Cell Signaling Technology, 9167; 1:1,000), and GAPDH (Sigma-Aldrich, G9545; 1:2,000).

Techniques: Infection, Luminex, Western Blot, Control, Expressing

Journal: PNAS Nexus

Article Title: Vascular endothelial cell-specific disruption of the profilin1 gene leads to severe multiorgan pathology and inflammation causing mortality

doi: 10.1093/pnasnexus/pgad305

Figure Lengend Snippet: Schematic representation of how endothelial Pfn1 loss could potentially lead to a proinflammatory condition. We propose that loss of Pfn1 in EC directly stimulates select proinflammatory cytokine/chemokine expression/release through upregulating the intrinsic IRF7/STAT1 signaling in addition to potentiating proinflammatory gene expression in monocytes/macrophages in a paracrine signaling manner.

Article Snippet: TCL was run on an SDS–PAGE and immunoblotted using antibodies specific for Pfn1 (Abcam, ab124904; 1:3,000), IRF7 (Cell Signaling Technology, Danvers MA, 72073; 1:1,000), p-IRF7 (Cell Signaling Technology, 24129; 1:1,000), STAT1 (Cell Signaling Technology, 9172; 1:1000), p-STAT1 (Cell Signaling Technology, 9167; 1:1,000), and GAPDH (Sigma-Aldrich, G9545; 1:2,000).

Techniques: Expressing, Gene Expression