Skip to main content
. Author manuscript; available in PMC: 2013 Dec 15.
Published in final edited form as: Cancer Res. 2012 Dec 7;72(24):6382–6392. doi: 10.1158/0008-5472.CAN-12-1033

Figure 3.

Figure 3

High TWIST1 expression allows for melanoma spheroid outgrowth. (A) Spheroids of WM793 cells constitutively expressing control or TWIST1 shRNAs were implanted into 3D collagen. Bright field images were taken at 24 hour increments imaging the same spheroids at each time point. Percent outgrowth was calculated by comparing the surface area of the spheroid (100%) to that of the total outgrowth at each time point. Columns, average of three independent experiments; bars, SD; **p<0.01. (B) WM35 constitutively overexpressing LacZ or TWIST1 was also assessed for spheroid outgrowth, as above. Columns, average of three independent experiments; bars, SD; **p <0.01. (C & D) At 96 hours, embedded spheroids were stained with both calcein AM (green = live cells) and ethidium bromide (red = dead cells) to determine apoptosis. One representative field of view is shown; scale, 100µm. (E & F) Alternatively, at 96 hours, embedded spheroids were subjected to EdU incorporation for 7 hours. Also, adherent cells grown in standard tissue culture dishes were subjected to EdU incorporation assay. Columns, average of three independent experiments; bars, SD.