Quantification of Cutaneous Ionocytes in Small Aquatic Organisms
[Abstract] Aquatic organisms have specialized cells called ionocytes that regulate the ionic composition, osmolarity, and acid/base status of internal fluids. In small aquatic organisms such as fishes in their early life stages, ionocytes are typically found on the cutaneous surface and their abundance can change to help cope with various metabolic and ...
Biofilm Assays on Fibrinogen-coated Silicone Catheters and 96-well Polystyrene Plates
[Abstract] Biofilm formation is a well-known bacterial strategy that protects cells from hostile environments. During infection, bacteria found in a biofilm community are less sensitive to antibiotics and to the immune response, often allowing them to colonize and persist in the host niche. Not surprisingly, biofilm formation on medical devices, such as ...
Measuring the Endocytic Recycling of Amyloid Precursor Protein (APP) in Neuro2a Cells
[Abstract] The established primary trigger of Alzheimer’s disease’s is β-amyloid (Aβ) (Mucke and Selkoe, 2012). Amyloid precursor protein (APP) endocytosis is required for Aβ generation at early endosomes (Rajendran and Annaert, 2012). APP retention at endosomes also depends on its recycling back to the plasma membrane (Koo et al., 1996; Ubelmann et ...
Indirect Immunofluorescence Assay in Chlamydomonas reinhardtii
Authors: Takashi Yamano and
Hideya Fukuzawa,
date: 07/05/2016,
view: 7066,
Q&A: 0
[Abstract] Determining the protein localization is essential to elucidate its in vivo function. Fluorescence-tagged proteins are widely used for it, but it is sometimes difficult to express tagged proteins in Chlamydomonas. Alternatively, indirect immunofluorescence assay is also one of the widely used methods and many reports determining ...
Immuno-EM Analysis of PF13_0191-GFP Expressing Parasites
[Abstract] This protocol was used to prepare pre-embedding samples of Plasmodium falciparum blood stage parasites that overexpressed the parasite protein PF13_0191 tagged with GFP. Using GFP-specific antibodies and Protein A-Gold the localisation of the overexpressed protein in the infected host cell was determined using standard transmission ...
Immunostaining Protocol: P-Smad2 (Xenograft and Mice)
Authors: Alexandre Calon,
Elisa Espinet,
Sergio Palomo-Ponce,
Daniele V. F. Tauriello,
Mar Iglesias,
María Virtudes Céspedes,
Marta Sevillano,
Cristina Nadal,
Peter Jung,
Xiang H.-F. Zhang,
Daniel Byrom,
Antoni Riera,
David Rossell,
Ramón Mangues,
Joan Massague,
Elena Sancho and
Eduard Batlle,
date: 05/05/2014,
view: 5059,
Q&A: 0
[Abstract] Metastasis depends on a gene program expressed by the tumor microenvironment upon TGF-beta stimulation. CRC (Colorectal cancer) cell lines did not induce robust stromal TGF-beta responses when injected into nude mice as shown by lack of p-SMAD2 accumulation in tumor-associated stromal cells. To enforce high TGF-beta signaling in xenografts, we ...
Immunostaining Protocol: P-Stat3 (Xenograft and Mice)
Authors: Alexandre Calon,
Elisa Espinet,
Sergio Palomo-Ponce,
Daniele V. F. Tauriello,
Mar Iglesias,
María Virtudes Céspedes,
Marta Sevillano,
Cristina Nadal,
Peter Jung,
Xiang H.-F. Zhang,
Daniel Byrom,
Antoni Riera,
David Rossell,
Ramón Mangues,
Joan Massague,
Elena Sancho and
Eduard Batlle,
date: 05/05/2014,
view: 6405,
Q&A: 0
[Abstract] We sought to understand the mechanisms behind the potent effect of stromal TGF-beta program on the capacity of colorectal cancer (CRC) cells to initiate metastasis. We discovered that mice subcutaneous tumors and metastases generated in the context of a TGF-beta activated microenvironment displayed prominent accumulation of p-STAT3 in CRC cells ...
α2β1-integrin Clustering and Internalization Protocol
[Abstract] α2β1-integrin clustering experiment can be used to trigger internalization of α2β1-integrin. When clustering is performed with sequential administration of primary and fluorescent secondary antibodies, the entry kinetics of integrin can be followed into the cell. The idea is first to allow binding of primary antibodies (recognizing the ...