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. 2013 Oct 3;8(10):e76247. doi: 10.1371/journal.pone.0076247

Figure 5. siRNA-mediated ENPEP, CK2-α, CCNJ, and MEGF9 knockdown recapitulates the biological effects of miR-125b expression.

Figure 5

(A) Western blot analysis of ENPEP, CK2-α, CCNJ, and MEGF9 upon transfection of the indicated siRNAs and scrambled control in MCF7 cells. Cells were transfected with the indicated pools of siRNAs at final concentrations of 30 nM by using HiPerFect transfection reagent. After 72 h of transfection, the cells were counted, photographed, and either collected for protein extraction or trypsinized for FACS analysis. (B) Cell numbers after 3 days of transfection with the indicated siRNAs. (C) Images of phenotypic effects on cell proliferation after 3 days of transfection with the indicated siRNAs in MCF7 cells. (D) The cell cycle profiles of MCF7 cells after expression with the indicated siRNAs or scrambled control. Scramble (G0/G1, 68.9%; S, 19.6%; G2/M, 11.7%), siCCNJ (G0/G1, 44.2%; S, 16.5%; G2/M, 39.2%), siMEGF9 (G0/G1, 53.2%; S, 18.5%; G2/M, 28.2%), siENPEP (G0/G1, 44.1%; S, 16.5%; G2/M, 39.3%), and siCK2-α (G0/G1, 43.8%; S, 22.3%; G2/M, 33.9%). Cells were transfected according to the conditions described in A. ***p < 0.001.