Supplementary Material for: Tissue-Specific Regulation of <b><i>Drosophila</i></b> NF-κB Pathway Activation by Peptidoglycan Recognition Protein SC

In <i>Drosophila</i>, peptidoglycan (PGN) is detected by PGN recognition proteins (PGRPs) that act as pattern recognition receptors. Some PGRPs such as PGRP-LB or PGRP-SCs are able to cleave PGN, therefore reducing the amount of immune elicitors and dampening immune deficiency (IMD) pathway activation. The precise role of PGRP-SC is less well defined because the <i>PGRP-SC</i> genes <i>(PGRP-SC1a</i>, <i>PGRP-SC1b</i> and <i>PGRP-SC2)</i> lie very close on the chromosome and have been studied using a deletion encompassing the three genes. By generating <i>PGRP-SC-</i>specific mutants, we reevaluated the roles of PGRP-LB, PGRP-SC1 and PGRP-SC2, respectively, during immune responses. We showed that these genes are expressed in different gut domains and that they follow distinct transcriptional regulation. Loss-of-function mutant analysis indicates that PGRP-LB is playing a major role in IMD pathway activation and bacterial load regulation in the gut, although PGRP-SCs are expressed at high levels in this organ. We also demonstrated that PGRP-SC2 is the main negative regulator of IMD pathway activation in the fat body. Accordingly, we showed that mutants for either <i>PGRP-LB</i> or <i>PGRP-SC2</i> displayed a distinct susceptibility to bacteria depending on the infection route. Lastly, we demonstrated that PGRP-SC1 and PGRP-SC2 are required in vivo for full Toll pathway activation by Gram-positive bacteria.