Journal: International Journal of Molecular Sciences

Article Title: Endothelial Cells Tissue-Specific Origins Affects Their Responsiveness to TGF-β2 during Endothelial-to-Mesenchymal Transition

doi: 10.3390/ijms20030458

Figure Lengend Snippet: Endothelial cells characterization regarding morphological, immunophenotyping, and vessel-like structures assay. ( A ) Phase contrast micrography demonstrating the polygonal morphology of aortic artery endothelial cells (PAEC), coronary artery endothelial cells (CAEC), human umbilical vein endothelial cells (HUVEC), and pulmonary artery endothelial cells (HPAEC) cells (100× magnification). ( B ) Immunophenotyping of ECs by flow cytometry. ( C ) All endothelial cells (PAEC, CAEC, HUVEC, and HPAEC) were able to form vessel-like structures when grown in matrigel, evidencing characteristics typical of CEs (40× and 100× magnification).

Article Snippet: We used distinct types of endothelial cells (ECs): CAEC (coronary artery endothelial cells, ATCC ® -Catalog No. PCS-100-020), PAEC (aortic artery endothelial cells, ATCC ® -Catalog No. PCS-100-011), HPAEC (pulmonary artery endothelial cells, ATCC ® -Catalog No. PCS-100-022) and HUVEC (human umbilical vein endothelial cells).

Techniques: Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Endothelial Cells Tissue-Specific Origins Affects Their Responsiveness to TGF-β2 during Endothelial-to-Mesenchymal Transition

doi: 10.3390/ijms20030458

Figure Lengend Snippet: Characterization of EndMT induction by TGF-β2 (10 ng/mL) in cell lines ( A ) PAEC, ( B ) CAEC, ( C ) HPAEC, and ( D ) HUVECs (non-treated or treated with TGF-β2). Immunofluorescence microscopy of cell lines induced to EndMT shows a decrease in the fluorescent intensity of CD31 (green) in PAECs, CAECs, and HUVECs cells. The nuclei were stained with DAPI (blue) and F-actin were stained with Phalloidin (red) (scale bar 50 µM; representative image of one replicate of each sample).

Article Snippet: We used distinct types of endothelial cells (ECs): CAEC (coronary artery endothelial cells, ATCC ® -Catalog No. PCS-100-020), PAEC (aortic artery endothelial cells, ATCC ® -Catalog No. PCS-100-011), HPAEC (pulmonary artery endothelial cells, ATCC ® -Catalog No. PCS-100-022) and HUVEC (human umbilical vein endothelial cells).

Techniques: Immunofluorescence, Microscopy, Staining

Journal: International Journal of Molecular Sciences

Article Title: Endothelial Cells Tissue-Specific Origins Affects Their Responsiveness to TGF-β2 during Endothelial-to-Mesenchymal Transition

doi: 10.3390/ijms20030458

Figure Lengend Snippet: TGF-β2 decrease formation of vessel-like structures in the cell lines (CAEC, PAEC, HPAEC, and HUVEC). The cells were treated with TGF-β2 and evaluated the capacity formation of vessel-like structures. This inhibitory effect was observed mainly in PAECs (representative image of one replicate; n = 3).

Article Snippet: We used distinct types of endothelial cells (ECs): CAEC (coronary artery endothelial cells, ATCC ® -Catalog No. PCS-100-020), PAEC (aortic artery endothelial cells, ATCC ® -Catalog No. PCS-100-011), HPAEC (pulmonary artery endothelial cells, ATCC ® -Catalog No. PCS-100-022) and HUVEC (human umbilical vein endothelial cells).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Endothelial Cells Tissue-Specific Origins Affects Their Responsiveness to TGF-β2 during Endothelial-to-Mesenchymal Transition

doi: 10.3390/ijms20030458

Figure Lengend Snippet: Effect of EndMT on the activation of the Erk pathway. The cells (CAEC, PAEC, HUVEC and HPAEC) were cultured for five days in presence TGF-β2 (10 ng/mL). Aliquots were withdrawn after the treatment and evaluated by ( A ) Multiplex technique analysis Array Kit ( n = 3, * p ≤ 0.05) and ( B ) western blotting using phospho-Erk1/2 (Thr202/Tyr204) and ERK1/2. β-actin were used as endogenous controls (representative image of one replicate of each sample). ( C ) Chemical inhibitor against MEK1/2 (U0126; 1 μM) inhibits the increase of ERK1/2 phosphorylation in the PAECs treated with TGF-β2. 1) U0126; 2) U0126-15′ TGF-β2; 3) U0126-30′ TGF-β2; 4) TGF-β2-15′; 5) TGF-β2-30′. GAPDH were used as endogenous controls (representative image of one replicate of each sample).

Article Snippet: We used distinct types of endothelial cells (ECs): CAEC (coronary artery endothelial cells, ATCC ® -Catalog No. PCS-100-020), PAEC (aortic artery endothelial cells, ATCC ® -Catalog No. PCS-100-011), HPAEC (pulmonary artery endothelial cells, ATCC ® -Catalog No. PCS-100-022) and HUVEC (human umbilical vein endothelial cells).

Techniques: Activation Assay, Cell Culture, Multiplex Assay, Western Blot, Phospho-proteomics

Journal: Journal of Cellular and Molecular Medicine

Article Title: High-mobility group box-1 induces vascular remodelling processes via c-Jun activation

doi: 10.1111/jcmm.12519

Figure Lengend Snippet: Human pulmonary arterial endothelial cells and smooth muscle cells express the HMGB1 receptors TLR4 and RAGE. Relative TLR4 and RAGE mRNA expression (A and B) and protein (C) expression in human pulmonary arterial endothelial cells (PAEC) and human pulmonary arterial smooth muscle cells (PASMC); D1–D3 represent samples from three different donors for PASMC or lots for PAEC.

Article Snippet: Primary human arterial endothelial cells (PAEC) were purchased from Lonza (Basel, Switzerland) ( n = 6), cultured in VascuLife® VEGF Medium (LifeLine Technology, Frederick, MD, USA) and used for experiments until passage 8.

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: High-mobility group box-1 induces vascular remodelling processes via c-Jun activation

doi: 10.1111/jcmm.12519

Figure Lengend Snippet: HMGB1-induced proliferation of pulmonary arterial endothelial cells and smooth muscle cells. Effect of HMGB1 treatment on human pulmonary arterial smooth muscle cells (PASMC, A–D) and human pulmonary arterial endothelial cells (PAEC, E–H) on (A and E) proliferation n = 5–6, (B and F) apoptosis n = 5, (C and G) attachment n = 5 and (D and H) migration n = 4–6. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Primary human arterial endothelial cells (PAEC) were purchased from Lonza (Basel, Switzerland) ( n = 6), cultured in VascuLife® VEGF Medium (LifeLine Technology, Frederick, MD, USA) and used for experiments until passage 8.

Techniques: Migration

Journal: Journal of Cellular and Molecular Medicine

Article Title: High-mobility group box-1 induces vascular remodelling processes via c-Jun activation

doi: 10.1111/jcmm.12519

Figure Lengend Snippet: HMGB1 activates MAPK intracellular signalling pathways. Western blot analysis for MAPK and downstream factor activation in (A) human pulmonary arterial smooth muscle cells (PASMC) and (B) human pulmonary arterial endothelial cells (PAEC) following stimulation with 1 or 100 ng/ml HMGB1 for the indicated time-points. Blots are representative of a minimum three independent experiments.

Article Snippet: Primary human arterial endothelial cells (PAEC) were purchased from Lonza (Basel, Switzerland) ( n = 6), cultured in VascuLife® VEGF Medium (LifeLine Technology, Frederick, MD, USA) and used for experiments until passage 8.

Techniques: Western Blot, Activation Assay

Journal: Frontiers in Genetics

Article Title: Mxi1-0 Promotes Hypoxic Pulmonary Hypertension Via ERK/c-Myc-dependent Proliferation of Arterial Smooth Muscle Cells

doi: 10.3389/fgene.2022.810157

Figure Lengend Snippet: Hypoxia induces expression of Mxi1-0 but not Mxi1-1 in PASMCs. (A) PASMCs were exposed to hypoxia (1% O 2 ) for indicated periods of time. Cell lysates were prepared and subjected to Western blotting assay. (B) Pulmonary arterial endothelial cells (PAECs) and PASMCs were exposed to normoxia (21% O 2 ) or hypoxia for 12 h, and were subjected to Western blottinganalyses. (C,D) PASMCs were transfected with constructs for HA-tagged Mxi1-0 (C) or Flag-tagged Mxi1-1 (D) , and were subjected to Western blotting analyses. (E) PASMCs were exposed to normoxia or hypoxia for 12 h, and were subjected to immunostaining with a Mxi1 antibody and counterstaining of the nuclei with DAPI. Scale bar, 20 μm. Data from three independent experiments are shown as means ± SDs. For statistical significance, *** represents p < 0.001 compared to normoxia or the mock-transfected group.

Article Snippet: Primary human PAECs were purchased from ScienCell, Inc (#3100) and cultured in endothelial cell medium (ECM, ScienCell).

Techniques: Expressing, Western Blot, Transfection, Construct, Immunostaining