Journal: Vascular Biology

Article Title: OxLDL induces the release of IL-1β from primed human endothelial and smooth muscle cells via different caspase -1-dependent mechanisms

doi: 10.1530/VB-22-0009

Figure Lengend Snippet: (A, B, C and D) Representative bright-field microscope images of stimulated, untreated HCAECs (A) and HCASMCs (C) compared with cytokine-stimulated (10 ng/mL of TNF-α and IL-1α for 48 h) HCAECs (B) and HCASMCs (D) treated with 50 µg/mL oxLDL for 6 h (HCAECs) or 2h (HCASMCs) and stained with Oil red O. Droplets stained with Oil red O were observed within both cell types upon stimulation and treatment. Images were taken using a Leica© bright-field microscope and are representative of 3 independent experiments, scale bar = 100 µm. (E and F) ELISA analysis of intracellular IL-1β in HCAECs (E) and HCASMCs (F) showed a significant increase following cytokine priming but no effect with oxLDL treatment when compared to the cytokine-only control. Data are presented as mean ± s.e.m. and analysed using a one-way ANOVA followed by a Bonferroni post-hoc test, **** P < 0.0001, *** P < 0.001, n = 3 independent donors.

Article Snippet: Human coronary artery endothelial (HCAECs) and smooth muscle cells (HCASMCs) were purchased from PromoCell (Heidelberg, Germany) and maintained according to the manufacturer’s instructions.

Techniques: Microscopy, Staining, Enzyme-linked Immunosorbent Assay

Journal: Vascular Biology

Article Title: OxLDL induces the release of IL-1β from primed human endothelial and smooth muscle cells via different caspase -1-dependent mechanisms

doi: 10.1530/VB-22-0009

Figure Lengend Snippet: (A and B) Cytokine stimulation and treatment with higher concentrations of oxLDL for 6 h (HCAECs) or 2 h (HCASMCs) induced significant IL-1β release from both HCAECs (A) and HCASMCs (B) compared to the cytokine-only control. (C and D) Increased release of IL-1β following cytokine-priming and treatment with 50 µg/mL oxLDL is associated with elevated LDH release in HCAECs (C) but not in HCASMCs (D). Samples are compared to the cytokine-only controls. Data are presented as mean ± s.e.m. and analysed using a one-way ANOVA followed by a Bonferroni post-hoc test, **** P < 0.0001, *** P < 0.001, n = 3 independent donors.

Article Snippet: Human coronary artery endothelial (HCAECs) and smooth muscle cells (HCASMCs) were purchased from PromoCell (Heidelberg, Germany) and maintained according to the manufacturer’s instructions.

Techniques:

Journal: Vascular Biology

Article Title: OxLDL induces the release of IL-1β from primed human endothelial and smooth muscle cells via different caspase -1-dependent mechanisms

doi: 10.1530/VB-22-0009

Figure Lengend Snippet: (A and B) ELISA analysis showed a significant reduction in oxLDL-induced IL-1β release from HCAECs following inhibition of both caspase-1 (YVAD) (A) and the NLRP3 inflammasome (MCC950) (B). (C and D) The release of IL-1β from oxLDL-treated HCASMCs was only significantly reduced upon inhibition of caspase-1 (C) and not the NLRP3 inflammasome (D). Data are expressed as mean ± s.e.m. and analysed using a one-way ANOVA and a Bonferroni post-hoc test, **** P < 0.0001, n = 3 independent donors.

Article Snippet: Human coronary artery endothelial (HCAECs) and smooth muscle cells (HCASMCs) were purchased from PromoCell (Heidelberg, Germany) and maintained according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Inhibition

Journal: Vascular Biology

Article Title: OxLDL induces the release of IL-1β from primed human endothelial and smooth muscle cells via different caspase -1-dependent mechanisms

doi: 10.1530/VB-22-0009

Figure Lengend Snippet: (A and B) ELISA analysis showed a significant reduction in oxLDL-induced IL-1β release from HCAECs following inhibition of gasdermin D (disulfiram) (A) but disulfiram was without effect on the release of IL-1β from HCASMCs (B). (C and D) The release of LDH was measured in HCAEC after cytokine treatment which was significantly reduced by 1 µM disulfiram treatment (C). There was no LDH release under any condition from HCASMC. Data are expressed as mean ± s.e.m. and analysed using a one-way ANOVA and a Bonferroni post-hoc test, *** P < 0.001, ** P < 0.01, n = 3 independent donors.

Article Snippet: Human coronary artery endothelial (HCAECs) and smooth muscle cells (HCASMCs) were purchased from PromoCell (Heidelberg, Germany) and maintained according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Inhibition

Journal: Nature medicine

Article Title: Atheroprotective roles of smooth muscle cell phenotypic modulation and the TCF21 disease gene as revealed by single-cell analysis

doi: 10.1038/s41591-019-0512-5

Figure Lengend Snippet: ( a ) Linkage disequilibrium relationships (LD, R 2 measure) of all genome-wide significant CAD-associated SNPs at the 6q23.2 locus (bottom), relative to the position of TCF21 and long non-coding RNAs (LINC01312 and TARID) within the locus (top). The R 2 color indicates the degree of LD between each pair of SNPs, and ranges from 0 (grey) to 1 (red). The corresponding R 2 values are also shown in each box. ( b ) Relationship between the number of genome-wide significant CAD risk alleles in each haplotype (x-axis) and TCF21 expression (y-axis) in 52 primary human coronary artery smooth muscle cell (HCASMC) lines. ( c ) Correlation between the magnitude of CAD risk imparted by each risk allele (x-axis) with relative TCF21 expression from that allele (y-axis) in 36 CAD-associated SNPS at the 6q23.2 locus in aortic tissue from the STARNET database. p-value was calculated using Pearson’s moment correlation coefficient. Grey shaded areas indicate 95% confidence intervals are based on Fisher’s Z-transform.

Article Snippet: A total of 65 primary human coronary artery smooth muscle cell (HCASMC) lines were purchased from PromoCell (catalog #C-12511), Cell Applications (catalog # 350-05a), Lonza (catalog #CC-2583), Lifeline Cell Technology (catalog #FC-0031) and ATCC (catalog #PCS-100-021).

Techniques: Genome Wide, Expressing