Fig. 2

PHF14 silencing in glioma cell lines decelerates proliferation and clonogenic growth. a, b The knockdown efficacy of PHF14 examined in U251, U87MG and A172 cells by qRT-PCR (a) and Western blot (b). c, d Cellular DNA replication is mitigated after PHF14 knocked down. Ten thousand U87MG, U251 and A172 cells per well were seeded in 96-well plates overnight. EdU and Hoechest co-staining was performed on the second day to assess cellular DNA replication activities (c) (Scale bar = 100 μm). Values are counted as EdU + cells/total cells (d). e–g PHF14 silenced cells showed weaker proliferation and colony formation potential. 500 U251 and A172 cells per well were cultured in 6-well plates in Complete growth medium for 10 days and then stained by 1% crystal violet for colony formation assay (e) and single colony morphology (f) (Scale bar = 200 μm). The number of colonies containing over 50 cells were counted up for analysis (g). h Cell colonies were stained by Phalloidin-cy3 after 10 days of culture for assessment of actin filaments containment and cellular protrusions (Scale bar = 50 μm) (*P < 0.05; **P = 0.01; ***P < 0.001; ****P < 0.0001)