Volume: 6 Issue: 17 —BIO-PROTOCOL

Biochemistry

Protein Expression, Purification and Crystallization of the Sxl-Unr-msl2 Ribonucleoprotein Complex Authors: Janosch Hennig and Michael Sattler, date: 09/05/2016, view: 6307, Q&A: 0

This protocol describes the expression, purification and crystallization of a ternary protein-protein-RNA complex, consisting of the two RNA recognition motifs (RRMs) of Sex-lethal (Sxl), the first of five cold shock domains of Upstream-of-N-Ras (Unr), and an 18-nucleotide region of msl2 mRNA, called the F fragment (Hennig et al ...

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Identification of RNA-binding Proteins Authors: Kazuya Masuda and Tadamitsu Kishimoto, date: 09/05/2016, view: 9311, Q&A: 0

This protocol describes the extraction of RNA-binding proteins (RBPs) from cell lysates. In order to pull down target RBPs, 5-bromo-UTP (BrUTP)-incorporated RNA probes are used, which are generated by in vitro transcription. The schematic diagram (Flowchart) with procedure is indicated (Figure1 and Figure 2).

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Measurement of Transferrin- and Non-transferrin-bound Iron Uptake by Mouse Tissues Authors: Supak Jenkitkasemwong, Chia-Yu Wang and Mitchell D. Knutson, date: 09/05/2016, view: 5755, Q&A: 0

Iron in blood plasma is bound to its transport protein transferrin, which delivers iron to most tissues. In iron overload and certain pathological conditions, the carrying capacity of transferrin can become exceeded, giving rise to non-transferrin-bound iron, which is taken up preferentially by the liver, kidney, pancreas, and heart. The ...

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Cell Biology

BODIPY 493/503 Staining of Neutral Lipid Droplets for Microscopy and Quantification by Flow Cytometry Authors: Bo Qiu and M. Celeste Simon, date: 09/05/2016, view: 37888, Q&A: 0

Lipid droplets (LDs) are ubiquitous, dynamic organelles and function as a storage depot for neutral lipids, including triglycerides and cholesterol esters (Walther and Farese, 2012). The movement of lipid species into and out of LDs impacts a variety of cellular processes, such as energy homeostasis, lipid-based signaling, and membrane homeostasis ...

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Developmental Biology

Olfactory Bulb (OB) Transplants Authors: Maria Savvaki and Domna Karagogeos, date: 09/05/2016, view: 5336, Q&A: 0

Transplantation in mouse brain slices is a powerful tool in order to study axon targeting and migrational events during development. Taking advantage of donors and recipients belonging to different genotypes, this technique allows researchers to assess the contribution of donor and/or recipient tissue by performing various combinations and to ...

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Immunology

Isolation of Joint-infiltrating Cells Authors: Aoi Akitsu and Yoichiro Iwakura, date: 09/05/2016, view: 7137, Q&A: 0

Infiltration of leukocytes into joints is one of the main features of autoimmune inflammatory arthritis. Here, we describe the protocol for isolation of joint-infiltrating cells in mice. This protocol is useful to analyze cell surface antigens and intracellular cytokines by flow cytometry.

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Microbiology

In vitro Cell Wall Stress Assay for Fusarium oxysporum Authors: Elena Pérez-Nadales and Antonio Di Pietro, date: 09/05/2016, view: 6851, Q&A: 0

In this protocol we describe a cell wall stress assay for the fungal pathogen F. oxysporum, based on exposure to the two anionic dyes Calcofluor White (CFW) and Congo Red (CR). Both compounds have been used to exert stress upon the fungal cell wall in vitro (Perez-Nadales and Di Pietro, 2015; Perez-Nadales and Di Pietro, 2011; ...

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Neuroscience

Analysis of Enteric Neural Crest Cell Migration Using Heterotopic Grafts of Embryonic Guts Authors: Rodolphe Soret and Nicolas Pilon, date: 09/05/2016, view: 5319, Q&A: 0

Hirschsprung disease (HSCR), also named aganglionic megacolon, is a severe congenital malformation characterized by a lack of enteric nervous system (ENS) in the terminal regions of the bowel (Bergeron et al., 2013). As the ENS notably regulates motility in the whole gastrointestinal track, the segment without neurons remains tonically ...

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Object-context Recognition Memory Test for Mice Authors: Sofia Kanatsou and Harm Krugers, date: 09/05/2016, view: 6490, Q&A: 0

The object in-context (OIC) task is a variant of the widely used object recognition (OR) task (Dix and Aggleton, 1999). The OIC task makes use of the fact that rodents have a natural tendency to explore novel environments and objects. The hippocampus appears to play a major role in the OIC task (much more so than in the original OR task), where ...

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Plant Science

Measurements of Proline and Malondialdehyde Content and Antioxidant Enzyme Activities in Leaves of Drought Stressed Cotton Authors: Tianzi Chen and Baolong Zhang, date: 09/05/2016, view: 15195, Q&A: 3

Drought stress negatively affects cotton plant growth and induces various biochemical and physiological responses in cotton plants. Proline content and antioxidant enzymes are thought to be associated with maintaining the structure of cellular components or with protecting cellular function. Study of cotton plant responses towards drought stress ...

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Simple Methods for Screening and Statistical Analysis of Leaf Epidermal Cells in Dicotyledonous Plants Authors: Jian-Jun Tao, Shou-Yi Chen and Jin-Song Zhang, date: 09/05/2016, view: 7911, Q&A: 0

Leaf epidermal cell size and number are positively correlated to leaf area. Stomata are specialized epidermal cells vital for gas exchange and water transpiration. So, observation and statistical analysis of the leaf epidermal cells are valuable for the study of leaf development and response to environmental stimulus. The classical method is using ...

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Salinity and Drought Treatment Assays in Kenaf (Hibiscus cannabinus L.) Authors: Xiaoping Niu, Meixia Chen, Aifen Tao and Jianmin Qi, date: 09/05/2016, view: 7043, Q&A: 0

Salinity and drought are the two main factors that cause fiber yield and quality losses in kenaf. It is reported that salinity and drought can affect more than 10% of arable land and cause a global decline in the average yields of major crops by more than 50%. Therefore, understanding plant tolerance of drought and salinity is of fundamental ...

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Cycloheximide Assays to Measure Protein Degradation in vivo in Plants Authors: Jonathan Gilkerson, Raymond Tam, Aimee Zhang, Kate Dreher and Judy Callis, date: 09/05/2016, view: 11777, Q&A: 0

The half-life of a protein is a characteristic property, and can be modulated by post-translational modifications, changes in subcellular localization, and/or interaction with other proteins or ligands. As one determinant of its steady-state level, a protein’s degradation represents an important distinguishing attribute relevant to its biological ...

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A Method to Analyze Local and Systemic Effects of Environmental Stimuli on Root Development in Plants Authors: Neil E. Robbins II and José Dinneny, date: 09/05/2016, view: 7089, Q&A: 0

Root development in vascular plants is innately tied to the environment. However, relatively little attention has been paid toward understanding the spatial scales at which the root perceives and responds to external stimuli. While some environmental signals elicit global responses that affect root system architecture, others may have more ...

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Stem Cell

Single-cell Visualization of Chromosome Transcriptional Territories by RNA-paint Authors: Céline Vallot and Claire Rougeulle, date: 09/05/2016, view: 6117, Q&A: 0

We developed a FISH-based method to directly assess chromosome-wide transcriptional activity, thereby enabling the visualization of the actively transcribed fraction of a chromosome at the single-cell level. We applied this method to probe the activity of X-chromosomes and its instability in the context of human embryonic stem cells and cancer ...

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Past Issues in 2016

Volume: 6 Issue: 17

Biochemistry

Cell Biology

Developmental Biology

Immunology

Microbiology

Neuroscience

Plant Science

Stem Cell