thapsigargin and aurin

Fluorescence in situ hybridization (FISH) and immunofluorescence (IF) are sensitive techniques used for detecting nucleic acids and proteins in cultured cells. However, these techniques are rarely applied together, and standard protocols are not readily compatible for sequential application on the same specimen. Here, we provide a user-friendly step-by-step protocol to perform multicolor RNA-FISH in combination with IF to simultaneously detect the subcellular localization of distinct RNAs and proteins in cultured cells. We demonstrate the use of our protocol by analyzing changes in the subcellular distribution of RNAs and proteins in cells exposed to a variety of stress conditions.

Topics: Antibodies; Arsenites; Aurintricarboxylic Acid; Base Sequence; Cytoplasmic Granules; DNA Helicases; Fluorescent Antibody Technique; Genes, Reporter; Green Fluorescent Proteins; HEK293 Cells; Hot Temperature; Humans; Hydrogen Peroxide; In Situ Hybridization, Fluorescence; Oligonucleotide Probes; Poly-ADP-Ribose Binding Proteins; Protein Binding; RNA Helicases; RNA Recognition Motif Proteins; RNA, Ribosomal, 18S; RNA, Ribosomal, 28S; Sodium Compounds; Stress, Physiological; Thapsigargin