Fig. 1From: Metalloprotease-disintegrin ADAM12 actively promotes the stem cell-like phenotype in claudin-low breast cancerADAM12 expression is upregulated in claudin-low breast cancer cells and in subpopulations enriched for CSCs. a. Cell surface expression of ADAM12 protein in breast cancer cell lines was evaluated by flow cytometry. Red, anti-ADAM12 antibody staining; gray, isotype control antibody staining. b Total cellular expression of ADAM12 was analyzed by Western blotting after partial purification of ADAM12 on concanavalin A (conA) agarose, as described [42]. GAPDH in the input fractions shows comparable conA agarose loading for all cells. Arrow, the nascent full length ADAM12; arrowhead, the processed form lacking the N-terminal pro-domain. c CSC signature score versus ADAM12 mRNA expression in 51 breast cancer cell lines. The CSC signature scores were calculated based on ref. [22] and microarray expression data retrieved from GEO:GSE69017, as described in Methods. Cell lines analyzed in panels a and b are shown in red. d ADAM12 protein levels in total lysates of SUM159PT cells grown as attached monolayers or as mammospheres, analyzed by Western blotting. Arrow, the nascent full length ADAM12; arrowhead, the processed form lacking the N-terminal pro-domain; *, a non-specific band. Positive control represents ADAM12 after partial purification on conA agarose. e Cell surface expression of ADAM12 in SUM159PT cells treated for 6Â days with DMSO (control) or with 10 nM paclitaxel (PTX), and then allowed to recover for 6Â days without PTX, was examined by flow cytometry. The population of cells with the highest expression of ADAM12 is shown in red. FSC, forward scatterBack to article page