SNF5 Is an Essential Executor of Epigenetic Regulation during Differentiation
Figure 3
Knockdown of SNF5 enhances a stem cell like state and blocks differentiation.
(A) SNF5, OCT4, EZH2, and loading control histone H3 were analyzed by western blot, 72 h post-transfection with SNF5 siRNA in NCCIT cells. (B and C) To get the nucleosome footprint, we have performed at least three biological replicates of NOMe-seq at 72 h post-transfection with SNF5 siRNA in NCCIT and selected ∼10 sequences in an unbiased manner to represented in the figures. (D) Stably infected SNF5 knockdown NCCIT cells were selected for 21 days with antibiotics and SNF5, OCT4, EZH2, and loading control histone H3 were subsequently analyzed by western blot. G401 cells were used for a SNF5 knockdown control. (E) At the same time point, cell morphology micrographs (200X) were taken. The data is representative of three biological experiments. (F) Apoptosis of SNF5 knockdown NCCIT cells after RA treatment was determined by flow cytometric analysis. The X axis indicates Annexin V and the Y axis indicates Propidium iodide (PI). The data are representative of three biological experiments (the mean +SEM).