Journal: bioRxiv

Article Title: Impaired SOX17 Expression Causes Endothelial Dysfunction and Pulmonary Arterial Hypertension by Insufficient Suppression of RUNX1

doi: 10.64898/2026.05.14.725187

Figure Lengend Snippet: (A) Experimental protocol for intervention of SuHx-PH in SOX17enhKO mice shows administration of the RUNX1 inhibitor Ro5-3335 or Ro24-7429 every other day for 6 times 1 week after the beginning of mild SuHx treatment. (B and C) RVSP (B) and RV/LV+S ratio (C) were measured at the end of week 3. Data in (B) and (C) are mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, ns: not significant.

Article Snippet: To generate stable RUNX1 overexpression HPAECs, full-length human RUNX1 cDNA (OriGene, Cat# SC123977) was cloned into the pHAGE-EF1α-IRES-blast vector from our lab. High titer lentiviruses were produced at Brown University Legorreta Cancer Center Cell & Genome Engineering Shared Resource.

Techniques:

Journal: bioRxiv

Article Title: Impaired SOX17 Expression Causes Endothelial Dysfunction and Pulmonary Arterial Hypertension by Insufficient Suppression of RUNX1

doi: 10.64898/2026.05.14.725187

Figure Lengend Snippet: (A) Representative Western blot analyses of endothelial and hematopoietic markers in control versus RUNX1 overexpression HPAECs, with quantification of relative band intensities normalized to loading controls. (B and C) Human Endothelial Cell Biology RT²-PCR Array analyses in HPAECs overexpressing RUNX1 compared to control vector transduced cells. Gene expression values were normalized to housekeeping genes and represented as fold change relative to controls. Bar graphs and Volcano plots highlight differentially expressed genes meeting the significance criteria of fold change > 1.5 and FDR < 0.05. *p < 0.05, **p < 0.01, ***p < 0.001 indicate statistical significance.

Article Snippet: To generate stable RUNX1 overexpression HPAECs, full-length human RUNX1 cDNA (OriGene, Cat# SC123977) was cloned into the pHAGE-EF1α-IRES-blast vector from our lab. High titer lentiviruses were produced at Brown University Legorreta Cancer Center Cell & Genome Engineering Shared Resource.

Techniques: Western Blot, Control, Over Expression, Reverse Transcription Polymerase Chain Reaction, Plasmid Preparation, Gene Expression

Journal: bioRxiv

Article Title: Impaired SOX17 Expression Causes Endothelial Dysfunction and Pulmonary Arterial Hypertension by Insufficient Suppression of RUNX1

doi: 10.64898/2026.05.14.725187

Figure Lengend Snippet: HPAECs transduced with RUNX1 overexpressing (RUNX1 OE) or control lentivirus were subjected to quantitative assays to evaluate RUNX1-driven phenotypic and functional changes. (A) Morphometric assessment (length/width ratio) demonstrated pronounced alterations in cell morphology in RUNX1 OE cells. (B) Tube formation assays demonstrated impaired angiogenic network assembly (C) Migration assays showed increased gap closure over time in RUNX1 OE cells. (D) MTS proliferation assays indicated enhanced growth in RUNX1 OE cells. (E) Reduced Caspase-3/7 activity following TNF-α treatment demonstrated decreased apoptosis in RUNX1 OE cells. All image-based analyses (A-C) were quantified using ImageJ software. Statistical significance for (D and E) was determined by two-way ANOVA with multiple comparisons, while all other comparisons were performed using unpaired two-tailed t-tests. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: To generate stable RUNX1 overexpression HPAECs, full-length human RUNX1 cDNA (OriGene, Cat# SC123977) was cloned into the pHAGE-EF1α-IRES-blast vector from our lab. High titer lentiviruses were produced at Brown University Legorreta Cancer Center Cell & Genome Engineering Shared Resource.

Techniques: Transduction, Control, Functional Assay, Migration, Activity Assay, Software, Two Tailed Test

Journal: bioRxiv

Article Title: Impaired SOX17 Expression Causes Endothelial Dysfunction and Pulmonary Arterial Hypertension by Insufficient Suppression of RUNX1

doi: 10.64898/2026.05.14.725187

Figure Lengend Snippet: Western blot analyses show (A) increased RUNX1 protein levels in SOX17 KO HPAECs, with quantification of relative band intensity normalized to β-ACTIN. (B and C) Treatment of SOX17 KO HPAECs with RUNX1 inhibitor Ro5-3335 at 0, 10 or 25 µM (B) or RUNX1-targeting siRNA (C) leads to partial restoration of endothelial marker expression in the absence of SOX17. *p < 0.05.

Article Snippet: To generate stable RUNX1 overexpression HPAECs, full-length human RUNX1 cDNA (OriGene, Cat# SC123977) was cloned into the pHAGE-EF1α-IRES-blast vector from our lab. High titer lentiviruses were produced at Brown University Legorreta Cancer Center Cell & Genome Engineering Shared Resource.

Techniques: Western Blot, Marker, Expressing

Journal: bioRxiv

Article Title: Impaired SOX17 Expression Causes Endothelial Dysfunction and Pulmonary Arterial Hypertension by Insufficient Suppression of RUNX1

doi: 10.64898/2026.05.14.725187

Figure Lengend Snippet: (A and B) RT-PCR analyses (A) and Western blot analyses (B) demonstrate increased RUNX1 expression and reduced endothelial marker levels in iPSC-ECs derived from SOX17 mutant patients compared with healthy controls. (C) Western blots shows in a dose-dependent manner RUNX1 inhibitor Ro5-3335 partially restored endothelial marker expression in iPSC-ECs derived from SOX17 mutant PAH patients. (D) Western blots shows RUNX1 targeting siRNA downregulation partially restored endothelial marker expression in iPSC-ECs derived from SOX17 mutant PAH patients. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 denote statistical significance.

Article Snippet: To generate stable RUNX1 overexpression HPAECs, full-length human RUNX1 cDNA (OriGene, Cat# SC123977) was cloned into the pHAGE-EF1α-IRES-blast vector from our lab. High titer lentiviruses were produced at Brown University Legorreta Cancer Center Cell & Genome Engineering Shared Resource.

Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing, Marker, Derivative Assay, Mutagenesis

Journal: bioRxiv

Article Title: Impaired SOX17 Expression Causes Endothelial Dysfunction and Pulmonary Arterial Hypertension by Insufficient Suppression of RUNX1

doi: 10.64898/2026.05.14.725187

Figure Lengend Snippet: RUNX1 expression in whole bone marrow (A) and whole lung (B) in SOX17enhKO and WT mice after 3 weeks of normoxia (Norm) or Sugen/hypoxia (SuHx). N = 5-15 mice per group, * P < 0.05, ** P < 0.01, ns: not significant.

Article Snippet: To generate stable RUNX1 overexpression HPAECs, full-length human RUNX1 cDNA (OriGene, Cat# SC123977) was cloned into the pHAGE-EF1α-IRES-blast vector from our lab. High titer lentiviruses were produced at Brown University Legorreta Cancer Center Cell & Genome Engineering Shared Resource.

Techniques: Expressing

Journal: bioRxiv

Article Title: Impaired SOX17 Expression Causes Endothelial Dysfunction and Pulmonary Arterial Hypertension by Insufficient Suppression of RUNX1

doi: 10.64898/2026.05.14.725187

Figure Lengend Snippet: Cdh5-CreERT2;Runx1(flox/flox);SOX17enhKO triple transgenic SRV mice were subjected to mild SuHx treatment. When the SRV mice were treated with corn oil without tamoxifen, they developed significantly elevated RVSP. When the SRV mice were treated with tamoxifen and placed under mild SuHx conditions, they exhibited normal RVSP. **** P < 0.0001.

Article Snippet: To generate stable RUNX1 overexpression HPAECs, full-length human RUNX1 cDNA (OriGene, Cat# SC123977) was cloned into the pHAGE-EF1α-IRES-blast vector from our lab. High titer lentiviruses were produced at Brown University Legorreta Cancer Center Cell & Genome Engineering Shared Resource.

Techniques: Transgenic Assay

Journal: bioRxiv

Article Title: Impaired SOX17 Expression Causes Endothelial Dysfunction and Pulmonary Arterial Hypertension by Insufficient Suppression of RUNX1

doi: 10.64898/2026.05.14.725187

Figure Lengend Snippet: Out of the 359 PAH patients, SOX17 expression was undetectable in 206 and detectable in 153 patients, whereas the expression of RUNX1 could be seen in all subjects. RUNX1 expression was significantly higher in patients with undetectable expression of SOX17 compared to those with detectable SOX17 expression. * P < 0.05.

Article Snippet: To generate stable RUNX1 overexpression HPAECs, full-length human RUNX1 cDNA (OriGene, Cat# SC123977) was cloned into the pHAGE-EF1α-IRES-blast vector from our lab. High titer lentiviruses were produced at Brown University Legorreta Cancer Center Cell & Genome Engineering Shared Resource.

Techniques: Expressing