Fig. 5

Copper chelation sensitises shCOMMD3 cells. A. Copper levels in both shCOMMD3 and shcontrol 4T07-TGL (tagged with TK-GFP-luciferase) cells evaluated using Agilent 7700 ICPMS instrument. The levels were expressed as Moles of Cu/Mg of protein. B. CF4 staining to reveal Cu + abundance in 4T1.2 and 66cl4 cells. The intensity per cell was calculated to reflect the level of Cu + abundance. C. Protein levels of ATP7A level in both shCOMMD3 and shcontrol 4T07-TGL (tagged with TK-GFP-luciferase) cells. Nuclei are stained with DAPI. D. Representative images of 4T07 shControl and shCommd3 acini were grown on GFR matrigel in 2% FCS in the presence or absence of 50 µM tetrathiomolybdate (TM). Acini size was quantified by measuring diameters on day 10; n = 2. E. Western blot analysis showing the effect of different concentrations of tetrathiomolybdate (TM) on PARP and ATP7A levels at 24 h. Valosin containing protein (VCP) used as a loading control