Figure 7.

Targeting CA IX reduces the transformation of TNBC cells in cancer stem-like cells. BT-549 (A) and MDA-MB-231 (B) cells were seeded in ultra-low attachment multiwall plates and grown in serum-free DMEM supplemented with B27, bFGF (20 ng/mL) and EGF (10 ng/mL) in normoxic and hypoxic conditions for seven days. TNBC cells grown in 1% O₂ were transfected with siRNA CA IX (100 nM) and siRNA Scr (100 nM) or treated with RC44 (100 µM) and grown as described above. Spheroid formation was analyzed under a phase-contrast microscopy and size and number of formed spheroids was calculated using ImageJ. Bars depict mean ± SD of three independent experiments. *** p < 0.0001; ** p < 0.001; * p < 0.01.