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. 2016 Apr 20;7(21):31177–31190. doi: 10.18632/oncotarget.8870

Figure 6. ZNF224 controls cell growth by regulating expressions of p21 and p53 via miR-663a.

Figure 6

A and B. MCF-7 cells were transfected with LNA-ctrl or miR-663a (20 nM) in the presence or absence of miR-663a antagonist (40 nM) as indicated. The expression of p21 and p53 was examined by qRT-PCR and immunoblot, respectively. GAPDH and tubulin were used as loading control in qRT-PCR and immunoblot, respectively. Data are from at least three independent experiments (n=3). C and D. In addition, colony forming ability was analyzed (*, vs. LNA-ctrl; **, vs. miR-663a) and subjected to iCelligence for the growth analysis in the presence or absence of CPT (0.1 μM). E and F. MCF-7 cells were transfected with FLAG-ZNF224 (2 μg) in the presence or absence of miR-663a antagonist (40 nM). The expression of p21, p53, miR-663a, and ZNF224 was examined by qRT-PCR and immunoblot, respectively. GAPDH and U6 RNA were used as loading control for mRNA and microRNA, respectively, in qRT-PCR, and tubulin was used as a loading control for immunoblot. Data are from at least three independent experiments (n=3). G and H. In addition, colony forming ability was analyzed (#, vs. EV; †, vs. ZNF224) and subjected to iCelligence for the growth analysis in the presence or absence of CPT (0.1 μM). I and J. MCF-7 cells were transfected with control or ZNF224 si-RNA (20 nM), and the expression of p21, p53, miR-663a, and ZNF224 was examined by qRT-PCR and immunoblot, respectively (**, vs. control si-RNA). GAPDH and U6 RNA were used as loading control for mRNA and microRNA, respectively, in qRT-PCR, and tubulin was used as a loading control for immunoblot. Data represent the mean ± SEM of three independent experiments. *P < 0.01, **P < 0.01, # P < 0.01, and † P < 0.01.