FIG 7.

KDM2A reduces rRNA transcription under mild starvation conditions in triple negative breast cancer (TNBC) cells. (A) Three days after transfection of control siRNA (control) or siRNA for KDM2A (siKDM2A or siKDM2A-w), MDA-MB-231 and MCF-7 cells were lysed and analyzed by Western blotting with antibodies to KDM2A and β-actin. (B) After the cells were prepared as described for panel A, RNA was isolated and analyzed by qRT-PCR with specific primers for KDM2A. The expression levels were normalized by TBP mRNA. (C) After the cells were prepared as described for panel A, KDM2A was detected by indirect immunofluorescence technique with anti-KDM2A (red) and antinucleolin (green) antibodies. (D) KDM2A reduces rRNA transcription under mild starvation conditions. After the cells were treated as described for panel A, MDA-MB-231 cells were cultured for 2 h in DMEM supplemented with 10% serum in the presence (+) or absence (−) of 3 mM 2DG. The amounts of pre-rRNA and KDM2A mRNA were measured by qRT-PCR. The expression levels were normalized by using polr2a mRNA.