Fig. 2: TSSC4 interacts with two U5 snRNA-containing complexes.
From: TSSC4 is a component of U5 snRNP that promotes tri-snRNP formation
a Complexes co-precipitating with TSSC4-FLAG were eluted by a FLAG peptide and resolved by ultracentrifugation on 10–30% glycerol gradient. RNA was isolated from each fraction, resolved on UREA-PAGE gels, and silver-stained (bottom). A whole-nuclear extract was run in parallel to identify fractions where individual snRNPs sediment (top). b After RNA preparation (a), proteins were isolated from the organic phase and detected by western blotting. Lighter complex sedimenting in fractions 14–16 contains primarily U5 snRNA and U5-specific proteins (PRPF8 and EFTUD2) and heavier complex migrating in fractions 19–20 contains U5 snRNA, U5-specific proteins, and the PRPF19 protein. Endogenous TSSC4 marked by an asterisk (*), TSSC4-FLAG marked by double asterisks (**). The positions of molecular weight markers are shown. a, b The experiment was performed two times with a similar result. Uncropped and unprocessed scans are provided in the Source Data file.
