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. 2017 May 22;8(43):73947–73963. doi: 10.18632/oncotarget.18166

Figure 4. Loss of HSF1 activity reduces growth of mammospheres and breast cancer stem cells.

Figure 4

(A) MCF7 and BT474 cells were seeded for the mammosphere assay as described in Materials and Methods. Spheres were collected and total protein was isolated. Sphere lysates and lysates from monolayer (ML) cells were subjected to immunoblotting using the indicated antibodies. (B) Cells were transfected with control or HSF1 siRNA followed by immunoblotting. (C–D) BT474 and MCF7 cells with and without HSF1 knockdown from (B) were seeded for the mammosphere assay. (E–F) BT474 and MCF7 cells were transfected with an empty vector, HSF1, or HSF1-S326A followed by seeding for the mammosphere assay. (G–H) BT474 and MCF7 cells were transfected with non-targeting siRNA + an empty vector, HSF1 siRNA + empty vector, or HSF1 siRNA + Slug followed by seeding for the mammosphere assay. (I–J) BT474 (G) and MCF7 (H) were treated with KRIBB11 (5 µM) for 24 hrs and then subjected to flow cytometry for the CD44highCD24lowESAhigh cell population and the percentage of cells in the population is reported. *Indicates significant difference (p < 0.05).