Figure 8

Knockdown of OCT4, NANOG, and ZIC1 in OCT4 -transduced breast cells (OTBCs) affects self-renewal. (a) Cell viability was assessed in OTBCs86-L1 cells upon transfection of short interfering RNAs (siRNAs) designed to knock down OCT4, NANOG, and ZIC1. Data were normalized to cells transfected with a mismatch siRNA pool. Mock refers to untransfected OTBCs86-L1 cells. Cells were transfected with the corresponding siRNAs and placed in self-renewal conditions for 72 hours. Data represent the mean ± standard deviation (SD) of three independent experiments and were analyzed with student t test with P values set at *P ≤0.01 and **P ≤0.001. (b) Knockdown of OCT4 in immortalized human mammary epithelial cells (HUMECs). Transfection of siRNAs was performed as described above. (c) Real-time polymerase chain reaction quantification of OCT4, NANOG, and ZIC1 mRNA expression after knockdown in OTBCs86-L1 cells; data were normalized to cells transfected with a mismatch siRNA pool. (d) Protein expression levels of OCT4 and ZIC1 assessed by Western blot in OTBCs86-L1 (MOCK), knockdown OTBCs86-L1 (OCT4 or ZIC1), and mismatch siRNA pool cell population (MISMATCH). One of three independent experiments is shown.