Harvested cells were washed with Dulbecco’s phosphate-buffered saline (dPBS), centrifuged at 125 × g for 5 min and then the flash-frozen pellets were stored at −80 °C. Extracted RNA (500 ng–2 µg) were used as templates to make cDNA libraries using a High Capacity RNA-to-cDNA kit (Applied Biosystems). TaqMan gene expression assays were designed using GAPDH (Hs03929097, VIC) as an endogenous control and CDH1 (Hs01023895_m1, FAM), GSTP1 (Hs00943350_g1, FAM), HSPA1A (Hs00359163_s1, FAM), MGMT (Hs01037698_m1, FAM), MLH1 (Hs00179866_m1, FAM), PKP3 (Hs00170887_m1, FAM), RHOXF2 (Hs00261259_m1, FAM), SOX17 (Hs00751752_s1, FAM), SRPX2 (Hs00997580_m1, FAM), or THBS2 (Hs01568063_m1, FAM) as targets (Thermo Fisher Scientific). Quantitative PCR (qPCR) was performed in 10 µL reactions by using TaqMan Universal Master Mix II with UNG and 2 μL of diluted cDNA from each sample (Applied Biosystems). Gene expression levels were calculated by “delta delta Ct” and normalized to control samples using ViiA7 version 1.2.2 or QuantStudio version 1.3 software (Applied Biosystems by Life technologies).
Thbs2
Manufactured by Thermo Fisher Scientific
THBS2 is a laboratory equipment product manufactured by Thermo Fisher Scientific. It is designed to support various experimental and analytical procedures in scientific research and testing environments. The core function of THBS2 is to provide a reliable and versatile platform for conducting targeted experiments and analyses, but a detailed description of its intended use is not available within the scope of this request.
Automatically generated - may contain errors
Lab products found in correlation
Gapdh, by Thermo Fisher Scientific (1 mentions)
Taqman universal master mix 2 with ung, by Thermo Fisher Scientific (1 mentions)
Quantstudio version 1, by Thermo Fisher Scientific (1 mentions)
High capacity rna to cdna kit, by Thermo Fisher Scientific (1 mentions)
Gstp1, by Thermo Fisher Scientific (1 mentions)
Hspa1a, by Thermo Fisher Scientific (1 mentions)
Lect2, by Abcam (1 mentions)
2 protocols using thbs2
1
Quantitative PCR Gene Expression Analysis
2
Histological Analysis of Aortic Atherosclerosis
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Aortic tissue with visible atherosclerotic lesion was selected, sampled, and submitted for histological evaluation according to American Heart Association (AHA) histological criteria [8 (link)]. During histological classification, lesions were designated by Roman numerals representing lesion grade. Clinically silent atherosclerotic plaques were included in this study, while ruptures and healed plaques (type VI) were not included.
In the present study, 32 aortic tissue samples were embedded in paraffin blocks. For paraffin sections, aortic specimens were fixed in 10% formalin for one day, processed through graded alcohols and xylene, and embedded in paraffin. Tissue sections were then stained with haematoxylin and eosin (H&E) or immunostained (Figure 1 ). The specimens were painted with CD68 for macrophage, and THBS-2 (Thermo Fisher Scientific, TSP) and LECT-2 (Abcam) antibodies for immunohistochemical staining.
THBS-2 immunohistochemical staining showed myocyte and endothelial immunoreactivity in LiMA tissue (Figure 1 ). However, aortic punches showed positive staining in myoctes. LECT-2 immunohistochemical staining of aortic tissue showed positive staining in myocte but LiMA tissue showed myocyte and endothelial immunoreactivity. Aortic and LiMA THBS-2 and LECT-2 levels were measured by semiquantitative immunoreactivity method.
In the present study, 32 aortic tissue samples were embedded in paraffin blocks. For paraffin sections, aortic specimens were fixed in 10% formalin for one day, processed through graded alcohols and xylene, and embedded in paraffin. Tissue sections were then stained with haematoxylin and eosin (H&E) or immunostained (
THBS-2 immunohistochemical staining showed myocyte and endothelial immunoreactivity in LiMA tissue (
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