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. 2015 Apr 1;10(4):e0120781. doi: 10.1371/journal.pone.0120781

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© 2015 Ponceau 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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Fig 4 — SrcY527F-MEF and HMEC-1 cells were transfected for 96 hr with different shRNA directed against αII-spectrin (Sp-shRNA) or an irrelevant shRNA (Nr-shRNA). (A and B) Western Blot analysis revealed a 60% decrease in αII-spectrin expression. Lamin A/C was used to control protein loading. Results revealed efficiency of shRNA3h and 4h (for HMEC-1), 1m and 3m (for SrcY527F-MEF) on αII-spectrin expression. (C) Invadosomes were counted 96 hr after cell transfection with shRNA (GFP positive cells): 300 transfected cells were counted from at least 3 independent experiments. The percentage of cells forming invadosomes was evaluated in control cells (Nr-shRNA) versus spectrin-depleted cells (Sp-shRNA). αII-Spectrin silencing significantly reduced the percentage of HMEC-1 and SrcY527F-MEF cells forming invadosomes.