Affinity purification of muscle nuclei from adult C. elegans. (A,B) Total nuclei after isolation, stained with DAPI (A) or showing mCherry fluorescence (B). (C–F) Affinity purification of nuclei from strains containing both NTF and birA transgenes (C,D) or only the NTF transgene (E,F). DAPI-stained nuclei (C,E) and autofluorescing beads (D,F) are shown. (G,H) Example of an affinity-purified nucleus from a strain containing both NTF and birA transgenes. mCherry fluorescence (G), anti-Flag staining (H), DAPI staining (I), and autofluorescing beads (J) are shown. (K) Western blots detecting the NTF in total and affinity-purified nuclei, probed with an anti-Flag antibody (top) and streptavidin (middle). Streptavidin also detects an endogenous biotinylated protein of ∼125 kDa that is present in all samples and serves as a loading control (asterisk). Western blot detecting histone H3 in affinity-purified nuclei (bottom).
