Immunocytochemical localization of RyRs in dissociated hippocampal neuronal cell cultures co-labeled with the F-actin label phalloidin
to identify actin-rich dendritic spines confirmed RyR immunoreactive puncta throughout the dendritic shafts of cultured hippocampal neurons (Figure 2B).
The control cells stained with rhodamine phalloidin
to visualize actin protein showed uniform staining pattern (figure 7a).
toxin may explain the first stage, with digestive disorders 24-48 h after ingestion.
Propranolol HCl was obtained from Acros Organics (Geel, Belgium), Lucifer Yellow CH dilithium salt, DAPI and EDTA from Sigma-Aldrich Chemie (Buchs, Switzerland), Verapamil HCl from Fluka Chemie (Buchs, Switzerland), TexasRed[R]-X Phalloidin
from Molecular Biotechnology (MoBiTec, Gottingen, Germany).
Cellular nuclei were stained with PI and cytoplasm was counterstained with Alexa Fluor 488 phalloidin
Louis, MO); F-actin was stained by rhodamine phalloidin
(1:40; Molecular Probes, Carlsbad, CA).
Cells were then fixed by paraformaldehyde, permeabilized and probed with antibody against [alpha]5 integrin subunit to test for changes in Fn receptor or with rhodamine phalloidin
to test for changes in actin or with antibody to vinculin to test for changes in focal contacts.
For actin, fixed cells were incubated with FITC conjugated phalloidin
The cells were then washed with PBS, and 400 [micro]l of fixative containing rhodamine phalloidin
(20 mM KP04, 10 mM Pipes, 5mM EGTA, 2 [micro]M MgCl2, 0.
Characterization of organic anion transporting polypeptide 1b2-null mice: essential role in hepatic uptake/toxicity of phalloidin
Actin cytoskeleton was visualized by staining with Alexa (488)-conjugated phalloidin
(1 U/sample, Invitrogen, Carlsbad, CA, USA) and cell nuclei were counterstained with propidium iodide (Sigma-Aldrich Corporation, Steinheim, Germany).
25% Triton X-100 for 2 min, and immersed in tetramethylrhodamine isothiocyanate (TRITC)-labeled phalloidin
(1:100 dilution; Sigma) in PBS for 20 min.