Nucleosome density shapes kilobase-scale regulation by a mammalian chromatin remodeler

SAMOSA, a method previously developed in the Ramani Lab, was employed to reveal nucleosome patterns from remodeling reactions in vitro with the Imitation Switch (ISWI) ATP-Dependent Remodeler ATPase, SNF2h. SAMOSA was further employed to reveal single-molecule nucleosome patterns in vivo from SNF2h knockout and re-expression murine embryonic stem cells. Investigating an ATP-Dependent Chromatin Remodeler ATPase via single-molecule chromatin profiling with m6A methyltransferase footprinting and SMRT long-read sequencing. 128 in vitro chromatin samples were analyzed (two replicates per condition), including the following controls: unremodeled methylated arrays and methylated/unmethylated unassembled template DNA. 35 in vivo samples were analyzed, including SAMOSA SNF2h KO/WT re-expression/E14 mESC SAMOSA samples (three biological replicates and technical replicates) and unmethylated/methylated mESC controls.