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. Author manuscript; available in PMC: 2012 Nov 23.
Published in final edited form as: Cell. 2011 Nov 23;147(5):1066–1079. doi: 10.1016/j.cell.2011.10.039

Figure 5. Zcchc11 inhibition blocks tumorigenicity and invasiveness of Lin28A-expressing breast cancer cells.

Figure 5

(A) q.RT-PCR analysis of Zcchc11 knockdown in MDA-MB-231 and T47D breast cancer cells. Error bars ±S.E.M. (n=3) (B) Inhibition of Zcchc11 expression does not affect let-7a expression in Lin28B-expressing cells (MDA-MB-231), while it up-regulates let-7a expression in Lin28A-expressing cells (T47D). Let-7a expression levels measured by q.RT-PCR in cells treated with siRNAs for 48hr. Error bars ±S.E.M. (n=3) (C) Inhibition of Zcchc11 expression did not affect the colony formation ability of MDA-MB-231 cells, but suppressed the colony formation ability of T47D cells. The number of colonies was evaluated 20 days post plating in soft agar. The experiment was repeated thrice and the statistical significance was calculated using Student’s t test. (D) Inhibition of Zcchc11 expression did not affect the invasiveness of MDA-MB-231 cells, but suppressed the invasive ability of T47D cells. The number of invasive cells was measured 16h post transfection with indicated siRNAs. In all assays, 10 fields per insert were scored and SD was measured. The experiment was repeated thrice and the statistical significance was calculated using Student’s t test. (E) Inhibition of Zcchc11 expression did not suppress tumor growth of MDA-MB-231 cells in xenografts, however it inhibited tumor growth of T47D cells in xenografts. The treatments with indicated siRNA were performed intraperitoneally (i.p.) for 5 cycles starting on day 15. Each treatment group consisted of 5 mice. (F) q.RT-PCR analysis of siRNA inhibition of Lin28B, Lin28A, and Zcchc11 in xenograft tumors (day 30) derived from MDA-MB-231 and T47D cells. Error bars ±S.E.M. (n=3). (G) Inhibition of Lin28B but not of Zcchc11 allows up-regulation of let-7a expression levels in MDA-MB-231 xenograft tumors (day 30). However, inhibition of Lin28A or Zcchc11 results in let-7a up-regulation in T47D xenograft tumors. Let-7a expression levels measured by q.RT-PCR on tumors untreated or treated with indicated siRNA. Error bars ±S.E.M. (n=3). See also Figure S3.