Suppression of ADF or cofilin expression affects invadopodia formation. A. Cells were cultured on fluorescent gelatin-coated cover slips and stained with fluorescent-phalloidin to visualize invadopodia (arrows). B and C. Representative Western blots for the expression of huADF.mRFP and huCofilin.mRFP in MTLn3 cells at 0, 24, 36, 48, 60 and 72 h after infection. Level of expression compared to endogenous ADF/cofilin was averaged from 3 or more blots. D. The degradation area of gelatin (μm2) was quantified in Metamorph and divided by number of cells in the same field and expressed as degradation area (μm2)/cell. n ≥ 30 cells in each experiment, three independent experiments. * p < 0.05, ** p < 0.01 versus control. Scale bar: 10 μm.NA, No Rescue Adenovirus; ADF.R, huADF.mRFP; ADF, huADF.RedTrack; COF.R; huCofilin.mRFP; COF, huCofilin.RedTrack.