ZNF300 Knockdown Inhibits Forced Megakaryocytic Differentiation by Phorbol and Erythrocytic Differentiation by Arabinofuranosyl Cytidine in K562 Cells
Figure 5
ZNF300 knockdown promotes proliferation in K562 cells.
(A) The same amount of control and ZNF300 knockdown cells (5×103) were plated in triplicates in a 24-well plate and the cell number was counted for consecutive 6 days. Data were statistics (mean ±SD) of representative results from 3 independent experiments with similar results. (B) Cell proliferation assay was also performed by using Cell Counting Kit-8. The absorbance at 450 nm was measured for consecutive 3 days and normalized to that of the first day. The cell proliferation was presented as relative absorbance. (C) Control and ZNF300 knockdown cells were fixed, permeablized, and stained with DAPI. The DNA content was analyzed by FACS. The distribution of cells in G0/G1, S, and G2/M phases was further analyzed by ModFit LT. Data were the statistics of representative results from 3 independent experiments with similar results. Numbers indicate the percentage (mean ±SD). (D) Bar graph of the statistics of cell cycle profiling experiments. (E) Cell lysates were prepared from control or ZNF300 knockdown cells and the protein expression level was detected by western blot with antibodies as indicated. HSC70 served as a protein loading control. ** indicates p<0.01. (F) The cytosol fraction (Cyto) and nucleus (Nucl) fraction of K562 cells treated with (+) or without (−) PMA were used for western blot with antibodies as indicated.