Fig 1: Levels of TPBG in the differentiating cells and effect of TPBG gene knockdown on differentiation of iPSCs into NSCs. (A) Daily changes in levels of TPBG as measured by DIA-MS. (B) Flow cytometry on day 7 for Nestin (left), TPBG (middle), and Nestin and TPBG (right). Blue: sample; gray: control. (C) Flow cytometric analysis of TPBG content in control cells (blue) and siRNA-treated cells (red) using an anti-TPBG antibody. Unstained cells are in gray. (D) Timeline of the knockdown experiment. On days 5 and 6 of NSC differentiation, TPBG was knocked down by siRNA, after which the cells were transferred to medium C and cultured for 14 days. Cells on day 21 of differentiation were stained with anti-Tuj1 antibody (green) and DAPI (blue). (E) Tuj1 in the cytoplasm wild-type (left), siRNA-negative control (middle), and siRNA-transfected (right) cells on day 21. Green: Tuj1; blue: 4′,6-diamidino-2-phenylindole (DAPI).