ADAM17 is not responsible for the meningococcus-induced EPCR shedding.

<p><b>(A)</b> ADAM17 membranous expression of HDMEC cells transfected with a siRNA against ADAM17 (blue) or control (green). Grey: isotype control. For the quantification of ADAM17 expression see supplemental data <b><a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1006981#ppat.1006981.s003" target="_blank">S3 Fig</a>. (B)</b> EPCR expression in HDMEC cells transfected with either a control siRNA (left) or ADAM17 siRNA (right) and infected with a WT <i>N</i>. <i>meningitidis</i> strain for 4 hours (red) or left non-infected (green). Grey: isotype control. For the quantification of EPCR expression see supplemental data <b><a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1006981#ppat.1006981.s003" target="_blank">S3 Fig</a>.</b> A Crispr/Cas9-mediated knockdown of ADAM17 expression was performed as described in Material and Methods in the Human Cerebral Microvessels Endothelial Cells line hCMEC/D3. <b>(C)</b> ADAM17 expression in the resulting cell line (blue) compared to that of the parental cell line (green). <b>(D)</b> EPCR expression in parental hCMEC/D3 cell line (left) or ADAM17-negative-hCMEC/D3 cell line (right) infected with a WT <i>N</i>. <i>meningitidis</i> strain for 4 hours (red) or left non-infected (green). The adhesion of <i>N</i>. <i>meningitidis</i> on hCMEC/D3 is reduced compared to the adhesion on HDMEC explaining the slighty reduced shedding phenotype observed with hCMEC/D3. Data are representative of 3 independent experiments. (grey: isotype control). For the quantification of EPCR expression see supplemental data <b><a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1006981#ppat.1006981.s005" target="_blank">S5 Fig</a>. (E)</b> Soluble EPCR in the supernatant of ADAM17-negative-hCMEC/D3 cells after infection with a WT strain or non-infected (control) cells. Data are mean (+/- SEM) of 3 independent experiments. *:<i>p</i> = 0.007 (t-test).