Fig 1: Tumor-derived sNKG2DLs are shed by ADAM10 and ADAM17. (A) Detection of sMICA and sULBP-2 levels by ELISA in serum samples from 35 patients with NB and 10 healthy controls. (B) MICA/B and ULBP1-3 expression in the NB cell lines were assessed using flow cytometry. (C) After 2 days of culture, sMICA and sULBP1-3 production in the culture supernatant was assessed using ELISA. (D) Cell surface expression of ADAM10 and ADAM17 in NB SH-SY-5Y cells was analyzed by flow cytometry. (E-G) SH-SY-5Y cells were pretreated with the ADAM10/ADAM17 promotor PMA, ADAM10 inhibitor GI–X, or ADAM17 inhibitor TAPI-1 for 24 h. Cell surface MFI of MICA/B and ULBP2 was detected by flow cytometry. The levels of sMICA and sULBP2 in the culture supernatant were analyzed by ELISA. Data are presented as the mean ± SEM. P-values were determined using a Mann-Whitney U test in (A). ****P<0.001. s, soluble; ADAM, a disintegrin and metalloproteinase; NB, neuroblastoma; MFI, mean fluorescence intensity.
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