Figure 4

Migration and invasion of knockdown and overexpression of ATF3 was investigated by wound closure assay and Boyden Chamber assay. (a) shows the distance of the migration fronts over 120 h set to T0 (start of experiment, 100%). Representative images of the migrating cells exhibited the total width of the wound at T0, 48 and 120 h for untreated and plasmid controls (pLOC, pGIPZ), ATF3 and shATF3 (b). Scale bar indicates 500 μm. Invasive capability of infected cells was quantified from Matrigel assays relative to controls set to 100% (±S.D. of three independent experiments) (c). Representative images of invading cells are shown in d for plasmid controls (pLOC, pGIPZ), ATF3 and shATF3. Scale bar indicates 500 μm, frames show ×2.5 magnification. Response to JS-K (up to 25 μM) by cells with knockdown and overexpression of ATF3 was measured by MTT assay (e). Viability of treated cells was plotted relative to viability of untreated controls set to 100% (±S.D. of three independent experiments). Relative cell proliferation of knockdown and overexpression was examined in BrdU incorporation assay over 72 h (f). Proliferation was normalized to T0 (start of experiment) set to 1. Asterisks (*P<0.05, **P<0.01, ***P<0.001) indicate significance to controls or T0.