Enrichment and Isolation of Cancer Stem Cells

BCC subsets were sorted as described (Patel et al, 2012 (link)). Briefly, BCCs, stably transfected with pEGFP1-Oct3/4, were selected, based on relative GFP expression using the FACSDiva (BD Biosciences). The top 5% GFP (Oct4^hi) cells contained CSCs (Patel et al, 2012 (link)). Further sorting was conducted for CSCs with specific knockdown, based on the fluorescence-tagged shRNA, or scramble vector: GFP^hi/RFP^hi: MDA-MB-231 Oct4 GFP/scramble RFP, MDA-MB-231 Oct4 GFP/CDH2-shRNA-RFP, and MDA-MB-231 Oct4 GFP/Cx43-shRNA-RFP cells were sorted on the FACSDiva (BD Biosciences) to select GFP^hi/RFP^hi cells.