Influence of GSK3 inhibition and beta Catenin stabilisation on the transcriptome of pre B cells

The goal of this study was to determine how GSK3 inhibition/beta Catenin stabilisation affects the transcription profile of pre B cells Overall design: B cell precursors were obtained from hematopoietic stem cell cultures of control or Mb1crex Catnb lox(ex3) mice as described above and sorted based on B220 expression by flow cytometry, after sorting cells were directly processed for RNA isolation. For experiments in which GSK3 was inhibited, B cell precursors were treated with LY2090314 over night and directly processed for RNA isolation without prior sorting. RNA was extracted using Quick-RNA MicroPrep (ZYMO RESEARCH) according to the manufacturer's instructions. The RNA concentration in RNAse free H2O was analyzed by measuring the absorption at 260 / 280 nm with a NanoDrop (Thermo Scientific/Peqlab). Additionally, unsorted wildtype B cell precursors were incubated with or without 40nM LY2090314 (Sigma) over night and RNA was isolated. RNA was snap frozen in liquid nitrogen, samples were processed and the transcriptome was analyzed by Novogene company.