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Cell Death in the Skin: How to Study Its Quality and Quantity?
The characterization of the quality and quantity of cell death has gained substantial interest over the past decades. More recently necroptosis as a programmed form of necrosis has been identified as an important additional form of cell death with relevance in the skin. Understanding how to assay cell death in specific is of critical importance for cancer research and treatment. Here we describe six different methods that can be used to assay cell viability and to study the quality or quantity of cultured human keratinocytes in vitro. These methods include crystal violet assay, hypodiploidy analysis, caspase-8 cleavage, re...
Source: Springer protocols feed by Molecular Medicine - December 19, 2012 Category: Molecular Biology Source Type: news

MicroRNA Profiling During Human Keratinocyte Differentiation Using a Quantitative Real-Time PCR Method
The terminal differentiation of epidermal keratinocytes requires transcriptional and posttranscriptional regulatory mechanisms. MicroRNAs (miRNAs) are small noncoding RNAs that play key roles during differentiation processes by regulating protein expression at the posttranscriptional level. Several studies have investigated miRNA expression in murine or human skin using northern blotting, microarrays, deep sequencing, or real-time PCR (Andl et al., Curr Biol 16:1041–1049, 2006; Hildebrand et al., J Invest Dermatol 131:20–29, 2011; Sonkoly et al., PLoS One 2:e610, 2007; Yi et al., Nat Genet 38:356–362, 200...
Source: Springer protocols feed by Molecular Medicine - December 19, 2012 Category: Molecular Biology Source Type: news

Global Proteome Analyses of SILAC-Labeled Skin Cells
Physiological functions of skin cells are often altered in diseases. Since the underlying molecular mechanisms are generally executed by proteins, it is of interest to assess protein dynamics in normal and pathologically altered cells. These can be readily analyzed in relevant cell culture models by quantitative mass spectrometry (MS)-based proteomics, which is the method of choice to track the concerted action and spatial relocation of unknown involved factors in an unbiased way. Different quantitative MS strategies have been used to characterize protein dynamics. In this chapter we describe in detail the use of stable is...
Source: Springer protocols feed by Molecular Medicine - December 19, 2012 Category: Molecular Biology Source Type: news