Journal: International Journal of Vascular Medicine

Article Title: Myosin VI and Associated Proteins Are Expressed in Human Macrophages but Do Not Play a Role in Foam Cell Formation in THP-1 Cells

doi: 10.1155/2013/516015

Figure Lengend Snippet: Primers, annealing temperatures, and product sizes for PCR. β 2 mg, β 2 microglobulin.

Article Snippet: Other antibodies including mouse polyclonal anti-AP-2 α 2 and rabbit polyclonal anti- β -microglobulin IgG (Abcam, Cambridge, UK), mouse polyclonal anti-GIPC1 IgG (Abnova, Heidelberg, Germany), rabbit polyclonal anti- β -actin (Cell Signaling Technology, Hitchin, UK), and horse radish peroxidase-conjugated goat anti-mouse and goat anti-rabbit IgG (Thermo Fisher Scientific, Cramlington, UK) were from various suppliers as indicated, and normal goat serum was from Dako UK Ltd (Ely, UK).

Techniques: Sequencing

Journal: International Journal of Vascular Medicine

Article Title: Myosin VI and Associated Proteins Are Expressed in Human Macrophages but Do Not Play a Role in Foam Cell Formation in THP-1 Cells

doi: 10.1155/2013/516015

Figure Lengend Snippet: Expression of mRNA for Myo6 and related proteins in THP-1 macrophages and HMDM. Total RNA was extracted from THP-1 macrophages (3 separate lysates) and HMDM (obtained from three individual donors). PCR was used to amplify gene products for Myo6 (a), Dab2 (b), AP-2 α 2 (c), and GIPC1 (d). Products were visualised on a 1.2% agarose gel containing 0.01% ethidium bromide (v : v).

Article Snippet: Other antibodies including mouse polyclonal anti-AP-2 α 2 and rabbit polyclonal anti- β -microglobulin IgG (Abcam, Cambridge, UK), mouse polyclonal anti-GIPC1 IgG (Abnova, Heidelberg, Germany), rabbit polyclonal anti- β -actin (Cell Signaling Technology, Hitchin, UK), and horse radish peroxidase-conjugated goat anti-mouse and goat anti-rabbit IgG (Thermo Fisher Scientific, Cramlington, UK) were from various suppliers as indicated, and normal goat serum was from Dako UK Ltd (Ely, UK).

Techniques: Expressing, Agarose Gel Electrophoresis

Journal: International Journal of Vascular Medicine

Article Title: Myosin VI and Associated Proteins Are Expressed in Human Macrophages but Do Not Play a Role in Foam Cell Formation in THP-1 Cells

doi: 10.1155/2013/516015

Figure Lengend Snippet: Effects of LDL and oxLDL on expression of mRNA and protein for Myo6 and related proteins in THP-1 macrophages. THP-1 cells were incubated with nLDL or oxLDL (50 μ g protein/mL) or an equal volume of PBS (control) for 8 h or 24 h, and total RNA or protein was isolated. The abundance of transcripts for (a) Myo6, (b) Dab2, (c) AP-2 α 2, and (d) GIPC1 was determined by qPCR (quantified using a standard curve and normalised using geNorm), and the expression of protein for (e) Myo6 and (f) Dab2 was assessed by immunoblotting using β -actin as the housekeeping gene (Con, control). Band density was quantified using Quantity One software (Biorad), and results for (g) Myo6 and (h) Dab2 are expressed as % control value. Data shown ((a)–(d), g, h) are the mean from 4 separate experiments and error bars show the SEM. Significance limits * P < 0.05, ** P < 0.01, *** P < 0.001 versus control; # P < 0.05, ## P < 0.01 versus nLDL (one-way ANOVA).

Article Snippet: Other antibodies including mouse polyclonal anti-AP-2 α 2 and rabbit polyclonal anti- β -microglobulin IgG (Abcam, Cambridge, UK), mouse polyclonal anti-GIPC1 IgG (Abnova, Heidelberg, Germany), rabbit polyclonal anti- β -actin (Cell Signaling Technology, Hitchin, UK), and horse radish peroxidase-conjugated goat anti-mouse and goat anti-rabbit IgG (Thermo Fisher Scientific, Cramlington, UK) were from various suppliers as indicated, and normal goat serum was from Dako UK Ltd (Ely, UK).

Techniques: Expressing, Incubation, Control, Isolation, Western Blot, Software

Journal: International Journal of Vascular Medicine

Article Title: Myosin VI and Associated Proteins Are Expressed in Human Macrophages but Do Not Play a Role in Foam Cell Formation in THP-1 Cells

doi: 10.1155/2013/516015

Figure Lengend Snippet: Inhibition of the expression of mRNA and protein for Myo6 and related proteins by siRNA. THP-1 macrophages were transfected with siRNA targeting Myo6, Dab2, AP-2 α 2, GIPC1 or a nonsilencing scrambled siRNA sequence (control) using HiPerFect transfection reagent. Cells were lysed at 24 h–96 h, and mRNA transcript levels for (a) Myo6, (b) Dab2, (c) AP-2 α 2, and (d) GIPC were determined by qPCR, and the expression of protein for (e) Myo6 and (f) Dab2 was assessed by immunoblotting. Band density was quantified using Quantity One software (Biorad), and results are expressed as % control value. +, siRNA; − scrambled siRNA. Data shown are the mean from 3 separate experiments, and error bars show the SEM. Significance limits, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 versus control; # P < 0.05, ## P < 0.01 versus 96 h (one-way ANOVA).

Article Snippet: Other antibodies including mouse polyclonal anti-AP-2 α 2 and rabbit polyclonal anti- β -microglobulin IgG (Abcam, Cambridge, UK), mouse polyclonal anti-GIPC1 IgG (Abnova, Heidelberg, Germany), rabbit polyclonal anti- β -actin (Cell Signaling Technology, Hitchin, UK), and horse radish peroxidase-conjugated goat anti-mouse and goat anti-rabbit IgG (Thermo Fisher Scientific, Cramlington, UK) were from various suppliers as indicated, and normal goat serum was from Dako UK Ltd (Ely, UK).

Techniques: Inhibition, Expressing, Transfection, Sequencing, Control, Western Blot, Software