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. 2014 Mar 21;9(3):e92687. doi: 10.1371/journal.pone.0092687

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© 2014 Fancsalszky et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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MDA-MB-231T cells were stably transfected with, pcDNA3 (K1 and K2), pcDNA3/FLAG-nm23-H1 (HA1 and HA2), pcDNA3/FLAG-ndk-1 (CE1 and CE2) and pcDNA3/MYC-nm23-H2 (HB1 and HB2). A: Western blot with anti-α-tubulin antibodies (loading control). B: Western blot with anti-FLAG- antibodies, visible band in HA1, HA2, CE1 and CE2 proves stable overexpression of introduced transgenes. C: Western blot with anti-MYC- antibodies, visible band in HB1 and HB2 (overexpression of NM23-H2). D: Migration assay. MDA-MB-231T cells stably transfected with one of the following constructs: pcDNA3 (K1 and K2), pcDNA3FLAG/nm23-H1 (HA1 and HA2), pcDNA3FLAG/ndk-1 (CE1 and CE2) and pcDNA3/MYC-nm23-H2 (HB1 and HB2) were tested for migration potential. The cells were stained with crystal violet and counted (the number of migrated cells were counted in four representative microscopic fields per each clone). The CE1 and CE2 clones as well as HA1, HA2, HB1 and HB2 exhibited significantly diminished migration potential compared to control (K1 and K2) clones (Student's t-test, p<0.05). The results are presented as an absolute number of migrated cells in 4 representative fields for every clone (±SD).