Figure 3
From: Computer-assisted quantification of motile and invasive capabilities of cancer cells
aZEcs, aMDIcs and aSDIcs quantifications reliably confirm manual measurements of HGF-induced cell dissemination in the presence of 10% serum.
(A) Representative images of 100× magnified zones of exclusion in monolayers of LA-EGFP expressing DAOY or UW228 cells in 96 well plate. Images are inverted greyscale of LA-EGFP fluorescence at 0 (T0h) and 24 h (T24h) −/+ stimulation with 20 ng/ml HGF and/or treatment with c-Met inhibitors PHA665752 and ARQ197 (125 nM each). (B,C) Means and SDs of % area covered from three independent zone infiltration experiments using aZEcs in DAOY (B) or UW228 (C) at T24h after HGF stimulation and/or inhibitor treatment. (D,E) Time-lapsed mean and SD quantifications of % area covered from three independent zone infiltration experiments using aZEcs in DAOY (D) or UW228 (E) after HGF stimulation and/or inhibitor treatment. (F) Representative images of 100x magnified microbeads coated with DAOY or UW228 cells after 24 h −/+ stimulation with 20 ng/ml HGF and/or treatment with c-Met inhibitors PHA665752 and ARQ197 (125 nM each). Inverted greyscale images of Hoechst-stained nuclei are shown. (G) Quantification of distance from the surfaces of the microbeads using aMDIcs. Box plots with pooled data of two independent experiments and whiskers min to max are shown. (H) Spheroid invasion assay with DAOY and UW228 cells. Treatment and imaging as in F), except that 50× magnification was used. (I) Quantification of distance from centres of spheroids using aSDIcs. Box plots with pooled data of two independent experiments and whiskers min to max are shown.
