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. Author manuscript; available in PMC: 2015 Oct 23.
Published in final edited form as: Oncogene. 2014 Jun 23;34(17):2178–2188. doi: 10.1038/onc.2014.177

Figure 8. Depletion of mTOR or HSF1 reduces mammosphere formation in vitro.

Figure 8

Cells were plated at low density in suspension culture (5000/well) on regular or low attachment tissue culture plates.

(A) Number of cells grown on regular tissue culture plates (left panels) and spheroids formed in low attachment plates (right panels) were calculated at day 12 after plating. mTOR control hairpin or shRNA, infected cells were plated at day zero. For HSF1 knockdown, doxycycline (2 ug / ml) was added to cultures every 3 days for continuous induction of the inducible HSF1 control hairpin or HSF1 shRNA expression. Experiments were repeated three times, reproducibly and mean values +/− standard deviation are indicated.

(B) Low magnification image of mammospheres formed by cells expressing control hairpins for mTOR experiment (Ctrl., upper panel), shRNA targeting mTOR (mTOR kd), tripZ ctrl for HSF1 experiment (Ctrl. lower panel) and the tripZ HSF1 knockdown MDAMB-231 cells (HSF1 kd)