University of Pretoria
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The analysis of diallyl trisulfide on cell viability, osteoclast formation, mitogen activated protein kinase protein expression, oxidative stress markers expression and accumulation of reactive oxygen species

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These datasets consist of the effects that diallyl trisulfide (DATS) has on osteoclast-forming RAW 264.7 murine macrophages. The resazurin data set shows how DATS affects the cell viability of RAW 264.7 murine macrophages. The resazurin data was analyzed using a microplate spectrophotometer provided by the University of Pretoria and quantified using GraphPad Prism 5 to form a bar graph. The tartrate-resistant acid phosphatase (TRAP) staining data was collected by manually counting and capturing (with an Olympus BH2 microscope) the number of osteoclasts in each well when exposed to varying concentrations of DATS. This data was quantified using GraphPad Prism 5 to form a bar graph. The western blot bands were analyzed using ImageJ software, which measured the protein band intensities of mitogen activated protein kinase and oxidative stress markers when exposed to DATS and RANKL. This data was quantified using GraphPad Prism 5 to form a bar graph.


The 2',7'-dichlorofluorescein diacetate (DCFDA) data was analyzed by manually counting and capturing 100 cells/well using a Zeiss Axiovert CFL40 microscope. A Zeiss Axiovert MRm monochrome camera and Zeiss filter 9 were operated to capture images of DCFDA-stained (green) cells. The fluorescence emission with at least 100 cells/well was assessed using ImageJ software in each experiment by analyzing the intensity of green fluorescence emitted in each cell. This data was then quantified using GraphPad Prism 5 to form a bar graph.

Funding

National Research Foundation (NRF), Grant Number: MND200607528963

History

Department/Unit

Physiology

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