Journal: International Journal of Molecular Sciences

Article Title: BMPR2 Loss Activates AKT by Disrupting DLL4/NOTCH1 and PPARγ Signaling in Pulmonary Arterial Hypertension

doi: 10.3390/ijms25105403

Figure Lengend Snippet: DLL4 is decreased in BMPR2-silenced PAECs and in lung tissue from patients with pulmonary arterial hypertension (PAH). Human primary pulmonary artery endothelial cells (PAECs) were transfected with control (siCTRL) or BMPR2 (siBMPR2) siRNA for 48 h and total protein lysates were collected for Western blotting of ( A ) DLL4, ( B ) N1ICD, ( C ) NOTCH1, ( D ) N2ICD, and ( E ) N4ICD. ( F ) DLL4 protein was also analyzed in PAECs transfected with either control (siCTRL) or BMPR2 (siBMPR2), CAV1 (siCAV1) or SMAD9 (siSMAD9) gene-specific siRNA pools. Representative Western blots are shown and densitometric analysis relative to β-actin and normalized to its corresponding siCTRL ( n = 5). ( G ) Immunohistochemical staining of DLL4 in paraffin embedded lung of failed donor controls (CTRL; n = 5), HPAH ( n = 3) and IPAH ( n = 2). Scale bar = 50 μm. Insert scale bar = 20 μm Data are presented as mean ± SEM; n = 5. ( A – E ) paired t -test, ( F ) 1-way ANOVA with Tukey HSD * p < 0.05; *** p < 0.005; and **** p < 0.001.

Article Snippet: Paraffin embedded lung tissue sections (10 μm) were cut (Histoserv, Inc., Germantown, MD, USA) and slides were stained for expression of DLL4 at the Lombardi Comprehensive Cancer Center Histopathology and Tissue Shared Resource (Georgetown University Medical Center, Washington, DC, USA).

Techniques: Transfection, Control, Western Blot, Immunohistochemical staining, Staining