primary human umbilical vein ecs huvecs  (ATCC)

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: TGF-β1 Promotes Angiogenesis via Endothelial-to-Mesenchymal Transition in Infantile Hemangioma

doi: 10.1161/ATVBAHA.125.322793

Figure Lengend Snippet: Changes in the expression of endothelial and mesenchymal markers after TGF-β1 (transforming growth factor β1) treatment in hemangioma-derived endothelial cells (HemECs) and human umbilical vein endothelial cells (HUVECs). A , Western blot analysis showing the expression of TGF-β1 in HemECs and HUVECs (n=4 biological replicates). The unpaired 2-tailed Student t tests were used. P =0.001, Cohen d =−7.50 (significant, extremely large effect). B , Immunofluorescence images showing the expression of endothelial and mesenchymal markers before and after TGF-β1 treatment in HemECs and HUVECs (n=4 biological replicates). Scale bar, 50 µm. C , Immunofluorescence analysis showing the mean fluorescence intensity of endothelial and mesenchymal markers before and after TGF-β1 treatment (n=4 biological replicates). The unpaired 2-tailed Student t tests were used. For marker expression in HemECs (TGF-β1 vs untreated), CD31: P =0.001, Cohen d =−7.14 (significant, extremely large effect). VE-cadherin (vascular endothelial cadherin): P =0.030, Cohen d =−2.93 (significant, very large effect). α-SMA (α-smooth muscle actin): P <0.001, Cohen d =13.55 (significant, extremely large effect). COL1A1 (collagen type I alpha 1 chain): P <0.001, Cohen d =11.58 (significant, extremely large effect). For marker expression in HUVECs (TGF-β1 vs untreated), CD31: P =0.72, Cohen d =−1.14 (not significant, large effect). VE-cadherin: P =0.21, Cohen d =1.65 (not significant, very large effect). α-SMA: P =0.92, Cohen d =−0.08 (not significant, negligible effect). COL1A1: P =0.93, Cohen d =−0.11 (not significant, negligible effect).

Article Snippet: HemEC isolation from proliferating IHs was performed as described previously., Primary human umbilical vein ECs (HUVECs) were obtained from the American Type Culture Collection (United States).

Techniques: Expressing, Derivative Assay, Western Blot, Immunofluorescence, Fluorescence, Marker

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: TGF-β1 Promotes Angiogenesis via Endothelial-to-Mesenchymal Transition in Infantile Hemangioma

doi: 10.1161/ATVBAHA.125.322793

Figure Lengend Snippet: TGF-β1 OE (TGF-β1 [transforming growth factor β1] overexpression) promotes hemangioma-derived endothelial cell (HemEC) migration, invasion, and angiogenesis. A , Transwell migration assay results showing the migration ability of HemECs and human umbilical vein endothelial cells (HUVECs) after TGF-β1 OE (n=4 biological replicates). Scale bar, 100 µm. The unpaired 2-tailed Student t tests were used. HUVECs: TGF-β1 OE vs control: P =0.96, Cohen d =0.44 (not significant, small to medium effect). HemECs: TGF-β1 OE vs control: P <0.001, Cohen d =8.85 (significant, extremely large effect). B , Transwell invasion assay results showing the invasion ability of HemECs and HUVECs after TGF-β1 OE (n=4 biological replicates). Scale bar, 100 µm. The unpaired 2-tailed Student t tests were used. HUVECs: P =0.62, Cohen d =−0.58 (not significant, medium effect). HemECs: P <0.001, Cohen d =9.29 (significant, extremely large effect). C , Tube formation assay showing the angiogenic ability of HemECs in vitro after TGF-β1 OE (n=4 biological replicates). Formed vessels are marked with yellow arrows. Scale bar, 100 µm. The unpaired 2-tailed Student t tests were used. For total tube length (TGF-β1 OE vs control), HUVECs: P =0.42, Cohen d =−0.79 (not significant, medium effect). HemECs: P <0.001, Cohen d =8.37 (significant, extremely large effect). For the number of branch sites (Control vs TGF-β1 OE ), HUVECs: P =0.60, Cohen d =−0.47 (not significant, medium effect). HemECs: P =0.017, Cohen d =3.19 (significant, extremely large effect). D , Vessel formation results showing the angiogenic ability of HemECs in vivo after TGF-β1 OE using 4 mice in each group (n=4). Scale bar, 50 µm. The unpaired 2-tailed Student t tests were used. TGF-β1 OE vs control: P =0.001, Cohen d =7.25 (significant, extremely large effect).

Article Snippet: HemEC isolation from proliferating IHs was performed as described previously., Primary human umbilical vein ECs (HUVECs) were obtained from the American Type Culture Collection (United States).

Techniques: Over Expression, Derivative Assay, Migration, Transwell Migration Assay, Control, Transwell Invasion Assay, Tube Formation Assay, In Vitro, In Vivo

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: TGF-β1 Promotes Angiogenesis via Endothelial-to-Mesenchymal Transition in Infantile Hemangioma

doi: 10.1161/ATVBAHA.125.322793

Figure Lengend Snippet: Changes in lipid metabolism. A , Oil Red O (ORO) staining showing the number of intracellular lipid droplets (LDs) in hemangioma-derived endothelial cells (HemECs) compared with that in human umbilical vein endothelial cells (HUVECs), with the etomoxir treatment group serving as a positive control (n=4 biological replicates). Scale bar, 20 µm. The Dunnett multiple comparisons test was used. HUVECs: TGF-β1 OE (transforming growth factor β1 overexpression) vs control: P =0.16, Cohen d =1.20 (not significant, large effect). Etomoxir vs control: P <0.001, Cohen d =10.54 (significant, extremely large effect). HemECs: TGF-β1 OE vs control: P =0.001, Cohen d =13.38 (significant, extremely large effect). Etomoxir vs control: P <0.001, Cohen d =24.63 (significant, extremely large effect). B , Western blot analysis showing that CPT1A (carnitine palmitoyltransferase 1A) protein expression decreased in HemECs (n=4 biological replicates). The unpaired 2-tailed Student t test was used. HemEC TGF-β1 OE vs control: P =0.001, Cohen d =−5.84 (significant, extremely large effect). C , Targeted metabolic analysis showing the top 30 differential metabolites (DMs) after TGF-β1 (transforming growth factor β1) treatment in HUVECs (n=6 biological replicates). D , Targeted metabolic analysis showing the top 30 differential metabolites (DMs) after TGF-β1 treatment in HemECs (n=6 biological replicates). E , Quantitative analysis of L-palmitoylcarnitine, with the etomoxir treatment group serving as a positive control (n=6 biological replicates). One-way ANOVA followed by the Dunnett multiple comparisons test was used to compare the TGF-β1 and etomoxir groups to the control group. HUVECs: TGF-β1 vs control: P =0.18, Cohen d =−0.83 (not significant, medium effect). Etomoxir vs control: P <0.001, Cohen d =−2.24 (significant, extremely large effect). HemECs: TGF-β1 vs control: P =0.003, Cohen d =−3.06 (significant, extremely large effect). Etomoxir vs control: P <0.001, Cohen d =−4.76 (significant, extremely large effect). F , Changes in the content of long-chain fatty acids (FAs; chain lengths C 14 –C 18 ; n=4 biological replicates). The paired 2-tailed Student t test with the Welch correction was performed separately for each FA chain length (C14, C16, and C18) to compare the TGF-β1–treated and control groups within HemECs and HUVECs. HemECs: C14: P =0.012, Cohen d =2.50 (significant, extremely large effect). C16: P =0.039, Cohen d =1.73 (significant, extremely large effect). C18: P =0.027, Cohen d =1.89 (significant, extremely large effect). HUVECs: C14: P =0.09, Cohen d =−1.37 (not significant, very large effect). C16: P =0.048, Cohen d =−1.60 (significant, large effect). C18: P =0.049, Cohen d =−1.64 (significant, extremely large effect). G , Palmitate-conjugated BSA (Palm-BSA) stimulated oxygen consumption rate (OCR) in HUVECs (n=4 biological replicates). A paired 2-tailed Student t test was performed. Control: P =0.009, Cohen d =4.11 (significant, extremely large effect). TGF-β1: P =0.010, Cohen d =0.55 (significant, medium effect). Etomoxir: P =0.84, Cohen d = –0.08 (not significant, negligible effect). TGF-β1+etomoxir: P =0.22, Cohen d =1.24 (not significant, large to very large effect). BSA: TGF-β1 vs control: P =0.36, Cohen d =−0.70 (not significant, medium effect). Palm-BSA: TGF-β1 vs control: P =0.08, Cohen d =−4.43 (not significant, extremely large effect). Etomoxir vs control: P <0.001, Cohen d =−4.83 (significant, extremely large effect). TGF-β1+etomoxir vs control: P =0.003, Cohen d =−3.30 (significant, extremely large effect). H , Palm-BSA failed to stimulate the OCR, thus inhibiting the fatty acid oxidation (FAO) effect of TGF-β1 or etomoxir treatment in HemECs (n=4 biological replicates). A paired 2-tailed Student t test was performed. Control: P =0.015, Cohen d =1.73 (significant, extremely large effect). TGF-β1: P =0.23, Cohen d =0.33 (not significant, medium effect). Etomoxir: P =0.64, Cohen d = –0.16 (not significant, small effect). TGF-β1+etomoxir: P =0.76, Cohen d =0.24 (not significant, negligible effect). BSA: TGF-β1 vs control: P =0.006, Cohen d =−2.97 (significant, extremely large effect). Palm-BSA: TGF-β1 vs control: P <0.001, Cohen d =−10.95 (significant, extremely large effect). Etomoxir vs control: P =0.002, Cohen d =−3.93 (significant, extremely large effect). TGF-β1+etomoxir vs control: P =0.003, Cohen d =−3.03 (significant, extremely large effect). I , Rate of FAO after TGF-β1 treatment in HUVECs and HemECs (n=4 biological replicates). The paired 2-tailed Student t test was performed. HemECs: P =0.038, Cohen d =−1.75 (significant, large effect). HUVECs: P =0.89, Cohen d =−0.08 (not significant, negligible effect).

Article Snippet: HemEC isolation from proliferating IHs was performed as described previously., Primary human umbilical vein ECs (HUVECs) were obtained from the American Type Culture Collection (United States).

Techniques: Staining, Derivative Assay, Positive Control, Over Expression, Control, Western Blot, Expressing

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: TGF-β1 Promotes Angiogenesis via Endothelial-to-Mesenchymal Transition in Infantile Hemangioma

doi: 10.1161/ATVBAHA.125.322793

Figure Lengend Snippet: CPT1A KD (CPT1A [carnitine palmitoyltransferase 1A] knockdown) stimulates hemangioma-derived endothelial cell (HemEC) migration, invasion, and angiogenesis. A , Transwell migration assay results showing the migration ability of HemECs and human umbilical vein endothelial cells (HUVECs) after CPT1A KD , CPT1A KD +L-palmitoylcarnitine, and CPT1A OE +TGF-β1 OE (TGF-β1 [transforming growth factor β1] overexpression; n=4 biological replicates). Scale bar, 100 µm. The unpaired 2-tailed Student t tests were performed. P values were adjusted using the Benjamini-Hochberg correction. HUVECs: CPT1A KD vs control: P =0.10, Cohen d =1.95 (not significant, large effect). CPT1A KD +L-palmitoylcarnitine vs CPT1A KD : P =0.038, Cohen d =−2.75 (significant, extremely large effect). TGF-β1 OE +CPT1A OE vs CPT1A KD +L-palmitoylcarnitine: P =0.10, Cohen d =1.74 (not significant, large effect). HemECs: CPT1A KD vs control: P <0.001, Cohen d =18.11 (significant, extremely large effect). CPT1A KD +L-palmitoylcarnitine vs CPT1A KD : P <0.001, Cohen d =−14.42 (significant, extremely large effect). TGF-β1 OE +CPT1A OE vs CPT1A KD +L-palmitoylcarnitine: P =0.13, Cohen d =−1.53 (not significant, large effect). B , Transwell invasion assay results showing the invasion ability of HemECs and HUVECs after CPT1A KD , CPT1A KD +L-palmitoylcarnitine, or CPT1A OE +TGF-β1 OE (n=4 biological replicates). Scale bar, 100 µm. The unpaired 2-tailed Student t tests were performed. P values were adjusted using the Benjamini-Hochberg correction. HUVECs: CPT1A KD vs control: P =0.19, Cohen d =1.29 (not significant, extremely large effect). CPT1A KD +L-palmitoylcarnitine vs CPT1A KD : P =0.99, Cohen d =0 (not significant, no effect). TGF-β1 OE +CPT1A OE vs CPT1A KD +L-palmitoylcarnitine: P =0.83, Cohen d =0.19 (not significant, small effect). HemECs: CPT1A KD vs control: P =0.005, Cohen d =4.30 (significant, extremely large effect). CPT1A KD +L-palmitoylcarnitine vs CPT1A KD : P =0.005, Cohen d =−4.31 (significant, extremely large effect). TGF-β1 OE +CPT1A OE vs CPT1A KD +L-palmitoylcarnitine: P =0.16, Cohen d =1.33 (not significant, large effect). C , Tube formation assay showing the angiogenic ability of HemECs in vitro after CPT1A KD , CPT1A KD +L-palmitoylcarnitine, and CPT1A OE +TGF-β1 OE (n=4 biological replicates). Scale bar, 100 µm. D , Vessel formation results showing the angiogenic ability of HemECs in vivo after CPT1A KD , CPT1A KD +L-palmitoylcarnitine, and CPT1A OE +TGF-β1 OE (n=4 biological replicates). Scale bar, 50 µm. The unpaired 2-tailed Student t tests were performed. P values were adjusted using the Benjamini-Hochberg correction. Total length: HUVECs: CPT1A KD vs control: P =0.90, Cohen d =0.11 (not significant, small effect). CPT1A KD +L-palmitoylcarnitine vs CPT1A KD : P =0.18, Cohen d =1.31 (not significant, large effect). TGF-β1 OE +CPT1A OE vs CPT1A KD +L-palmitoylcarnitine: P =0.32, Cohen d =−0.93 (not significant, large effect). HemECs: CPT1A KD vs control: P =0.003, Cohen d =2.73 (significant, extremely large effect). CPT1A KD +L-palmitoylcarnitine vs CPT1A KD : P =0.013, Cohen d =−3.53 (significant, extremely large effect). TGF-β1 OE +CPT1A OE vs CPT1A KD +L-palmitoylcarnitine: P =0.84, Cohen d =0.17 (not significant, small effect). Number of branch sites. HUVECs: CPT1A KD vs control: P =0.91, Cohen d =0.10 (not significant, small effect). CPT1A KD +L-palmitoylcarnitine vs CPT1A KD : P =0.71, Cohen d= −0.33 (not significant, medium effect). TGF-β1 OE +CPT1A OE vs CPT1A KD +L-palmitoylcarnitine: P =0.85, Cohen d =0.16 (not significant, small effect). HemECs: CPT1A KD vs control: P =0.001, Cohen d =3.35 (significant, extremely large effect). CPT1A KD +L-palmitoylcarnitine vs CPT1A KD : P =0.013, Cohen d =−6.40 (significant, extremely large effect). TGF-β1 OE +CPT1A OE vs CPT1A KD +L-palmitoylcarnitine: P =0.66, Cohen d =−0.38 (not significant, small effect). D , Vessel formation results showing the angiogenic ability of HemECs in vivo after CPT1A KD , CPT1A KD +L-palmitoylcarnitine, and CPT1A OE +TGF-β1 OE (n=4 biological replicates). Scale bar, 50 µm. The unpaired 2-tailed Student t tests with the Welch correction were performed. CPT1A KD vs control: P <0.001, Cohen d =8.90 (significant, extremely large effect). CPT1A KD +L-palmitoylcarnitine vs CPT1A KD : P =0.014, Cohen d =−3.81 (significant, extremely large effect). TGF-β1 OE +CPT1A OE vs CPT1A KD +L-palmitoylcarnitine: P =0.72, Cohen d =0.25 (not significant, negligible to small effect).

Article Snippet: HemEC isolation from proliferating IHs was performed as described previously., Primary human umbilical vein ECs (HUVECs) were obtained from the American Type Culture Collection (United States).

Techniques: Knockdown, Derivative Assay, Migration, Transwell Migration Assay, Over Expression, Control, Transwell Invasion Assay, Tube Formation Assay, In Vitro, In Vivo

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: TGF-β1 Promotes Angiogenesis via Endothelial-to-Mesenchymal Transition in Infantile Hemangioma

doi: 10.1161/ATVBAHA.125.322793

Figure Lengend Snippet: CPT1A KD (CPT1A [carnitine palmitoyltransferase 1A] knockdown) suppresses hemangioma-derived endothelial cell (HemEC) autophagy. A , Transmission electron microscopy (TEM) results showing changes in autophagy after CPT1A KD , CPT1A KD +L-palmitoylcarnitine, and CPT1A OE +TGF-β1 OE (TGF-β1 [transforming growth factor β1] overexpression) in HemECs and human umbilical vein endothelial cells (HUVECs; n=4 biological replicates). Scale bar, 1 µm. B , Autophagic vesicles per field (n=4 biological replicates). The unpaired 2-tailed Student t tests were performed. P values were adjusted using the Benjamini-Hochberg correction. HUVECs: CPT1A KD vs control: P =0.33, Cohen d =0.83 (not significant, large effect). CPT1A KD +L-palmitoylcarnitine vs CPT1A KD : P =0.11, Cohen d =−1.54 (not significant, large effect). TGF-β1 OE +CPT1A OE vs CPT1A KD +L-palmitoylcarnitine: P =0.06, Cohen d =1.89 (not significant, large effect). HemECs: CPT1A KD vs control: P =0.004, Cohen d =−3.24 (significant, extremely large effect). CPT1A KD +L-palmitoylcarnitine vs CPT1A KD : P <0.001, Cohen d =9.71 (significant, extremely large effect). TGF-β1 OE +CPT1A OE vs CPT1A KD +L-palmitoylcarnitine: P =0.017, Cohen d =−2.55 (significant, large effect). C , Immunoblot analysis of TGF-β1, CPT1A, and AMPK (5’-monophosphate [AMP]–activated protein kinase; n=3 biological replicates).

Article Snippet: HemEC isolation from proliferating IHs was performed as described previously., Primary human umbilical vein ECs (HUVECs) were obtained from the American Type Culture Collection (United States).

Techniques: Knockdown, Derivative Assay, Transmission Assay, Electron Microscopy, Over Expression, Control, Western Blot

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: TGF-β1 Promotes Angiogenesis via Endothelial-to-Mesenchymal Transition in Infantile Hemangioma

doi: 10.1161/ATVBAHA.125.322793

Figure Lengend Snippet: Changes in the expression of endothelial and mesenchymal markers after R(+) propranolol or S(−) propranolol treatment in hemangioma-derived endothelial cells (HemECs) and human umbilical vein endothelial cells (HUVECs). A , Immunofluorescence images showing the expression of endothelial and mesenchymal markers before and after R(+) propranolol or S(−) propranolol treatment in HemECs and HUVECs. Scale bar, 100 µm (n=4 biological replicates). B , Immunofluorescence analysis showing the mean fluorescence intensity of endothelial and mesenchymal markers before and after R(+) propranolol or S(−) propranolol treatment in HemECs (n=4 biological replicates). The Dunnett multiple comparisons test was used. R(+) propranolol vs control: CD31: P <0.001, Cohen d =16.23 (significant, extremely large effect). VE-cadherin (vascular endothelial cadherin): P <0.001, Cohen d =11.58 (significant, extremely large effect). α-SMA (α-smooth muscle actin): P <0.001, Cohen d =−20.50 (significant, extremely large effect). COL1A1 (collagen type I alpha 1 chain): P =0.001, Cohen d =−11.23 (significant, extremely large effect). S(−) propranolol vs control: CD31: P =0.20, Cohen d =0.99 (not significant, large effect). VE-cadherin: P =0.07, Cohen d =2.62 (not significant, very large effect). α-SMA: P =0.75, Cohen d =0.21 (not significant, small effect). COL1A1: P =0.88, Cohen d =0.11 (not significant, negligible effect). C , Immunofluorescence analysis showing the mean fluorescence intensity of endothelial and mesenchymal markers before and after R(+) propranolol or S(−) propranolol treatment in HUVECs (n=4 biological replicates). The Dunnett multiple comparisons test was used. R(+) propranolol vs control: CD31: P =0.46, Cohen d =0.54 (not significant, medium effect). VE-cadherin: P =0.24, Cohen d =−0.94 (not significant, large effect). α-SMA: P =0.65, Cohen d =0.33 (not significant, small effect). COL1A1: P =0.52, Cohen d =0.48 (not significant, small effect). S(−) propranolol vs control: CD31: P =0.10, Cohen d =−1.36 (not significant, very large effect). VE-cadherin: P =0.79, Cohen d =1.90 (not significant, very large effect). α-SMA: P =0.93, Cohen d =−0.07 (not significant, negligible effect). COL1A1: P =0.87, Cohen d =0.1 (not significant, negligible effect).

Article Snippet: HemEC isolation from proliferating IHs was performed as described previously., Primary human umbilical vein ECs (HUVECs) were obtained from the American Type Culture Collection (United States).

Techniques: Expressing, Derivative Assay, Immunofluorescence, Fluorescence, Control

Journal: Cardiovascular Research

Article Title: FLT4 causes developmental disorders of the cardiovascular and lymphovascular systems via pleiotropic molecular mechanisms

doi: 10.1093/cvr/cvae104

Figure Lengend Snippet: Subcellular localization and proteostatic signalling consequences of FLT4 TOF variants. ( A ) HUVECs expressing WT, MD, and TOF FLT4-V5 variants, stained with anti-V5 (red, FLT4), phalloidin (green, actin filaments), and DAPI (blue, nuclear stain). Scale bar, 10 μm. ( B ) Cells scored for three types of V5 staining, perinuclear/ER, PM/cytoplasmic, or both. One hundred cells in each group scored in each of three biological repeats. ( C ) Subcellular fractionation followed by immunoblotting for COS7 cells expressing FLT4 WT, MD, TOF-DNV, or two TOF-PTV variants. Fractions: A—PM, cytoplasmic; B—vesicular/Golgi apparatus-associated; or C—nuclear/perinuclear, ER. Markers: ACTB, cytoplasmic or nucleoplasmic cytoskeletal; GM130, Golgi apparatus; H3, histone 3, nuclear marker. V5-C-terminally tagged FLT4 variants. ( D ) Colocalization of FLT4-V5 tagged proteins with markers of the ER: calnexin; PM, Na/K-transporter; GM130, Golgi apparatus. ( E and F ) The activation of gene expression of proteostatic signalling by FLT4-TOF-DNV and FLT4-TOF-PTV variants. n = 3; ** P < 0.01; *** P < 0.001. ( G ) Activation of proteostatic signalling in HEK293T cells measured through HSP5A protein expression assessed by immunoblot. ( H ) Densiometric analysis of HSP5A bands relative to actin/EV from ( C ). n = 3, * P < 0.05; ** P < 0.01, **** P < 0.001; one-way ANOVA compared with EV.

Article Snippet: Primary human umbilical vein ECs (HUVECs) pooled from four donors (lot number: 420Z015.1) were obtained from Promocell (C-12203).

Techniques: Expressing, Staining, Fractionation, Western Blot, Marker, Activation Assay

Journal: Cardiovascular Research

Article Title: FLT4 causes developmental disorders of the cardiovascular and lymphovascular systems via pleiotropic molecular mechanisms

doi: 10.1093/cvr/cvae104

Figure Lengend Snippet: Hypoxia mimetics regulate the stability of an FLT4/ FLT4-TOF-PTV. RNA ( A ) or protein ( B ) was prepared from primary human ECs (HUVECs) expressing the FLT4 minigene assay construct (as outlined in , ). EV, WT, or Q736* PTV FLT4 minigenes were untreated, or treated with the hypoxia mimetics, 0.2 mM cobalt chloride (CC) or 100 nM DMOG, for 3 h. Red dagger, full-length minigene protein, both V5 and FLAG positive; blue dagger, truncated minigene protein caused by the introduction of a nonsense codon and concomitant C-terminal cleavage, V5 positive only; n = 3; ** P < 0.01; **** P < 0.0001.

Article Snippet: Primary human umbilical vein ECs (HUVECs) pooled from four donors (lot number: 420Z015.1) were obtained from Promocell (C-12203).

Techniques: Expressing, Mini Gene Assay, Construct

Journal: Cardiovascular Research

Article Title: FLT4 causes developmental disorders of the cardiovascular and lymphovascular systems via pleiotropic molecular mechanisms

doi: 10.1093/cvr/cvae104

Figure Lengend Snippet: Distinct transcriptomic profiles of primary human ECs expressing both types of TOF variants compared with WT or an MD FLT4. ( A ) Distribution of DEGs between HUVECs expressing FLT4-V5 WT, FLT4 R1041P (MD), and FLT4 TOF-DNV (C51W) or FLT4-TOF-PTV (1-736Q*), identified by RNAseq. ( B ) Reactome analysis of the 702 TOF FLT4-specific DEGs, up- or down-regulated examined separately, displaying pathways identified with FDR < 0.01. ( C ) Volcano plot showing the 702 FLT4 TOF-specific DEGs, with genes taken forward for qPCR analysis labelled, and those previously associated with CHD in red.

Article Snippet: Primary human umbilical vein ECs (HUVECs) pooled from four donors (lot number: 420Z015.1) were obtained from Promocell (C-12203).

Techniques: Expressing

Journal: Cardiovascular Research

Article Title: FLT4 causes developmental disorders of the cardiovascular and lymphovascular systems via pleiotropic molecular mechanisms

doi: 10.1093/cvr/cvae104

Figure Lengend Snippet: Rescue of FLT4 TOF-specific DEGs, represented as a heat map of gene expression levels, both ( A ) up-regulated and ( B ) down-regulated, by inhibitors of the three major proteostatic signalling pathways. Heat map of RNA levels (yellow highest, dark blue lowest) from HUVECs expressing WT FLT4-V5, MD, TOF-DNV, or TOF-PTV variants; the latter two treated with specific inhibitors of the three main proteostatic signalling pathways, or WT treated with the same inhibitors, above, IRE1α (red), PERK (blue), or ATF6 (green). Row names, gene targets. Red crosses represent no significant rescue ( P still <0.05 compared with WT) and orange crosses represent DEGs whose expression changes not recused by drug treatment ( P < 0.05 compared to WT still) of the gene expression changes induced by FLT4 TOF variant expression compared with WT.

Article Snippet: Primary human umbilical vein ECs (HUVECs) pooled from four donors (lot number: 420Z015.1) were obtained from Promocell (C-12203).

Techniques: Expressing, Variant Assay