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. 2012 Jun 7;33(10):1897–1908. doi: 10.1093/carcin/bgs198

Fig. 5.

Fig. 5.

3′-UTR-deficient PTEN or dominant negative Akt prevents miR-21-induced CSF-1 expression. MDA-MB-231 (panels A and C) and MCF-7 (panels B and D) breast cancer cells were cotransfected with CMV-miR-21 and 3′-UTR-less PTEN (panels A and B) or Akt K179M (panels C and D). Total RNAs were assayed for the presence of CSF-1 mRNA by qRT–PCR as described in Materials and methods. Mean ± SE of triplicate measurements is shown. *P < 0.001 versus control in panels A to D; **P < 0.001 versus CMV-miR-21-transfected for panels A to D. Bottom parts show PTEN and Akt K179M expression in cells transfected in parallel. (E to H) MDA-MB-231 (panels E and G) and MCF-7 (panels F and H) cells were transfected with CSF-1-Luc reporter along with 3′-UTR-less PTEN (panels E and F) or Akt K179M (panels G and H). The cell lysates were assayed for luciferase activity as described in Materials and methods (23,24,29,30). Mean ± SE of triplicate measurements is shown. *P < 0.01 versus control; **P < 0.01 versus CMV miR-21-transfected for panel E. *P < 0.05 versus control; **P < 0.01 versus CMV miR-21-transfected for panel F. *P < 0.05 versus control; **P < 0.05 versus CMV miR-21-transfected for panel G. *P < 0.01 versus control; **P < 0.01 versus CMV miR-21-transfected for panel H. Bottom parts show HA-tagged PTEN and Akt K179M expression in cells transfected in parallel.