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. Author manuscript; available in PMC: 2018 Oct 17.
Published in final edited form as: Oncogene. 2018 Apr 17;37(28):3879–3893. doi: 10.1038/s41388-018-0239-7

Figure 2. miR-106b-25 increases TIC characteristics in-vitro and in-vivo in a Notch dependent manner.

Figure 2

(A) Flow cytometry analysis in MCF7 cells treated with 30nM of either non-targeting siRNAs or pooled siRNAs that target NOTCH1 and transiently transfected with either a negative mimic or a combination of miR-106b, miR-93, and miR-25 mimics (20 nM total) to assess the percentage of CD44+/CD24− cells. (B) Secondary mammosphere assay in MCF7- and Sum159-NS and Cluster (CLR) stable cell lines in the presence of DAPT (5μM) or a DMSO vehicle control. (C) Left: Secondary mammosphere assay in MCF7 NS and CLR stable cell lines with stable shRNA knock down of NOTCH1. Right: Primary mammosphere assay in Sum-159 NS and Cluster stable cell lines with stable shRNA knock down of NOTCH1. Statistical significance on all mammosphere assays was assessed by one-way ANOVA followed by a Tukey post-test on triplicate samples of a representative experiment (n=3). (D) MCF7-NS and CLR cells with stable shRNA KD of Notch1 or a scramble control (SCR) were transplanted into the 4th mammary fat pad of NOG/SCID mice at limiting dilutions, data represents presence of tumors at week 5 post tumor cell implantation.