Supplementary Material for: The <b><i>Drosophila</i></b> Deubiquitinating Enzyme dUSP36 Acts in the Hemocytes for Tolerance to <b><i>Listeria monocytogenes</i></b> Infections

<i>Listeria monocytogenes</i> is a facultative intracellular pathogen which can infect<i> Drosophila melanogaster</i>. Upon infection, <i>Drosophila</i> mounts an immune response including antimicrobial peptide production and autophagy activation. A set of previously published results prompted us to study the role of the deubiquitinating enzyme dUSP36 in response to <i>L. monocytogenes</i> infections. We show in this report that flies with <i>dUsp36</i>-specific inactivation in hemocytes are susceptible to <i>L. monocytogenes</i> infections (as are flies with autophagy-deficient hemocytes) but are still able to control bacterial growth. Interestingly, flies with <i>dUsp36</i>-depleted hemocytes are not sensitized to infection by other pathogens. We conclude that <i>dUsp36</i> plays a major role in hemocytes for tolerance to <i>L. monocytogenes</i>

Supplementary Material for: The Nonaspanins TM9SF2 and TM9SF4 Regulate the Plasma Membrane Localization and Signalling Activity of the Peptidoglycan Recognition Protein PGRP-LC in Drosophila

Transmembrane 9 (TM9) proteins, or nonaspanins, are a family of proteins conserved throughout evolution and characterized by 9 transmembrane domains. In <i>Drosophila</i>, TM9 superfamily protein member 4 (TM9SF4) and its closest paralogue, TM9SF2, contribute to phagocytosis of various types of particles, while TM9SF4 displays non-redundant requirement in Gram-negative bacteria engulfment. In addition, the two TM9 proteins control the actin cytoskeleton in larval haemocytes and in <i>Drosophila </i>S2 cells. Here, we show that TM9SF4 and TM9SF2 co-immunoprecipitate with the peptidoglycan recognition protein (PGRP)-LC, which triggers the <i>Drosophila</i> immune response to bacterial infection. Furthermore, both TM9 proteins co-localize with this receptor in intracellular vesicles and at the plasma membrane in <i>Drosophila</i> S2 cells in culture and in the fly fat body. Silencing <i>TM9SF4 </i>prevents plasma membrane localization of PGRP-LC, whereas silencing <i>TM9SF2 </i>does not, which may account for the non-redundant role of TM9SF4 in phagocytosis of Gram-negative bacteria. Finally, we provide a set of data suggesting that TM9 proteins can prevent inappropriate signalling from the unstimulated receptor