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. 2011 Feb 28;6(2):e17168. doi: 10.1371/journal.pone.0017168

Figure 2. PMA-induced HB-EGF shedding is ADAM17-dependent.

Figure 2

A) AP-HB-EGF release measured as alkaline phosphatase activity (OD 405 nm) in conditioned media and B) surface AP-HB-EGF measured as cell surface fluorescence intensity per cell (FLU/cell) in DMSO control-treated (C; black bars) and 30 min, 400 nM PMA-treated (PMA; white bars) AP-HB-EGF expressing HT1080 cells treated with the ADAM17 inhibitor TAPI-2 (10 µM) or vehicle control. C, D) as in A and B respectively, except that cells were reverse transfected with control (C) or ADAM17 siRNAs for 72 h before treatment. The insert shows western blot analysis of total cell extracts from cells used in C and D, demonstrating efficient siRNA-mediated knockdown of pro and mature forms (arrows) of ADAM17. α-tubulin is used as an internal loading control. All graphs show average values ± standard error of the mean of at least three independent experiments each done in triplicate. *p<0.05; **p<0.01, ***p<0.001 after one-way analysis of variance with Bonferroni's post tests for multiple comparisons. Unless otherwise indicated the comparison is relative to the respective control.