Journal: Cell & Bioscience

Article Title: Serum IP-10 and IL-17 from Kawasaki disease patients induce calcification-related genes and proteins in human coronary artery smooth muscle cells in vitro

doi: 10.1186/s13578-020-00400-8

Figure Lengend Snippet: IP-10/IL-17 secreted in the KD plasma initiates the calcification of HCASMCs. a HCASMCs were either used as controls or were treated with either FC or KD plasma for 7, 14, or 21 days; b the HCASMCs were either maintained as controls or were treated with IP-10, IL-17, or both, for 7, 14, or 21 days; c the HCASMCs were either maintained as controls or were treated with FC or KD plasma, or KD plasma co-mixed with IP-10 and IL-17 blocking antibodies, for 21 days; a – c the calcification of the HCASMCs was analyzed using ARS stain. All data were mean ± SEM from three independent experiments; * P < 0.05 vs. control cells

Article Snippet: Human coronary artery SMCs (HCASMCs, ATCC ® PCS-100-021TM) were purchased from the cell bank (ATCC, Rockville, MD).

Techniques: Clinical Proteomics, Blocking Assay, Staining, Control

Journal: Cell & Bioscience

Article Title: Serum IP-10 and IL-17 from Kawasaki disease patients induce calcification-related genes and proteins in human coronary artery smooth muscle cells in vitro

doi: 10.1186/s13578-020-00400-8

Figure Lengend Snippet: KD plasma induces OPN, OCN, and ALP expressions in HCASMCs. The HCASMCs were either maintained as controls or were treated with either FC or KC plasma for 3, 7, or 10 days, after which we examined the mRNA expressions of OPN ( a ), OCN ( b ), and ALP ( c ) using real-time PCR and the protein expressions of OPN, OCN, and ALP ( d – e ) using the western blot test. The data in ( a – e ) are mean ± SEM from three independent experiments. * P < 0.05 vs. control cells. The results in ( d – e ) are representative of three independent experiments with similar results

Article Snippet: Human coronary artery SMCs (HCASMCs, ATCC ® PCS-100-021TM) were purchased from the cell bank (ATCC, Rockville, MD).

Techniques: Clinical Proteomics, Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: Cell & Bioscience

Article Title: Serum IP-10 and IL-17 from Kawasaki disease patients induce calcification-related genes and proteins in human coronary artery smooth muscle cells in vitro

doi: 10.1186/s13578-020-00400-8

Figure Lengend Snippet: IP-10/IL-17 co-treatment induces OPN, OCN and ALP expressions in HCASMCs. The HCASMCs were maintained either as controls or were treated with IP-10, IL-17, or both, for 3, 7, or 10 days, after which we examined the mRNA expressions of OPN ( a ), OCN ( b ), and ALP ( c ) using real-time PCR and the protein expressions of OPN, OCN, and ALP ( d – e ) using the western blot test. The data in ( a – e ) are mean ± SEM from three independent experiments. * P < 0.05 vs. control cells. The results in ( d – e ) are representative of three independent experiments with similar results

Article Snippet: Human coronary artery SMCs (HCASMCs, ATCC ® PCS-100-021TM) were purchased from the cell bank (ATCC, Rockville, MD).

Techniques: Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: Cell & Bioscience

Article Title: Serum IP-10 and IL-17 from Kawasaki disease patients induce calcification-related genes and proteins in human coronary artery smooth muscle cells in vitro

doi: 10.1186/s13578-020-00400-8

Figure Lengend Snippet: IP-10/IL-17 co-treatment-initiated HCASMC calcification is mediated by the BMP6 autocrine effect. a The HCASMCs were either maintained as the control or were co-treated with IP-10 and IL-17 for 3 days, after which we examined the BMP2/4/6 mRNA expression in the HCASMCs; b – d the HCASMCs were either maintained as the control or were pre-treated with IgG, BMP2-, BMP4-, or BMP6-blocking antibodies and then co-treated with IP-10 and IL-17; b HCASMC calcification was analyzed using ARS stain; c we examined the mRNA expressions OPN, OCN, and ALP using real-time PCR; d the protein expressions of OPN, OCN, and ALP were examined using the western blot test; e the HCASMCs were either maintained as the control or were treated with IP-10, IL-17, or both, for 48, 72, or 96 h, after which we examined the smad1/5 phosphorylation using the western blot test; f the HCASMCs were pretreated with control-, smad1- or smad5-specific siRNA and then either maintained as the control or co-treated with IP-10 and IL-17 for 14 days. The calcification of the HCASMCs was analyzed using ARS stain. The data in ( a – f ) are mean ± SEM from three independent experiments. * P < 0.05 vs. control cells; # P < 0.05 vs. IgG/or si-CL/IP-10 and IL-17-co-treated cells. The results in ( d – e ) are representative of three independent experiments with similar results

Article Snippet: Human coronary artery SMCs (HCASMCs, ATCC ® PCS-100-021TM) were purchased from the cell bank (ATCC, Rockville, MD).

Techniques: Control, Expressing, Blocking Assay, Staining, Real-time Polymerase Chain Reaction, Western Blot, Phospho-proteomics

Journal: Cell & Bioscience

Article Title: Serum IP-10 and IL-17 from Kawasaki disease patients induce calcification-related genes and proteins in human coronary artery smooth muscle cells in vitro

doi: 10.1186/s13578-020-00400-8

Figure Lengend Snippet: Runx2 regulates the BMP6 autocrine effect on the IP-10/IL-17 co-treatment-initiated OPN/OCN/ALP expression and calcification of HCASMCs. a The HCASMCs were either maintained as the control or were co-treated with IP-10 and IL-17 for 2, 4, 6, and 8 days; b the HCASMCs were either maintained as the control or were pre-treated with IgG or BMP6-blocking antibodies and then co-treated with IP-10 and IL-17 for 8 days; c the HCASMCs were pretreated with smad1- and smad5-specific siRNA and then either maintained as the control or co-treated with IP-10 and IL-17 for 8 days; a – c the expression of runx2 was examined using the western blot test; d – e the HCASMCs were pretreated with control- or runx2-specific siRNA and then maintained as either the control or co-treated with IP-10 and IL-17; d we examined the mRNA expressions of OPN, OCN, and ALP through real-time PCR; e the calcification of the HCASMCs was analyzed using ARS stain. The results in ( a – e ) are representative of three independent experiments with similar results. The data in ( d – e ) are mean ± SEM from three independent experiments. * P < 0.05 vs. siCL/control cells; # P < 0.05 vs. siCL/IP-10 and IL-17-co-treated cells

Article Snippet: Human coronary artery SMCs (HCASMCs, ATCC ® PCS-100-021TM) were purchased from the cell bank (ATCC, Rockville, MD).

Techniques: Expressing, Control, Blocking Assay, Western Blot, Real-time Polymerase Chain Reaction, Staining

Journal: Cell & Bioscience

Article Title: Serum IP-10 and IL-17 from Kawasaki disease patients induce calcification-related genes and proteins in human coronary artery smooth muscle cells in vitro

doi: 10.1186/s13578-020-00400-8

Figure Lengend Snippet: Schematic representation of the signaling pathways regulating KD plasma circulating IP-10 and IL-17 co-treatment-initiated vascular calcification in HCASMCs

Article Snippet: Human coronary artery SMCs (HCASMCs, ATCC ® PCS-100-021TM) were purchased from the cell bank (ATCC, Rockville, MD).

Techniques: Protein-Protein interactions, Clinical Proteomics