Matsayapan Pudla1, Panthong Kulsantiwong2, Chanya Srisaowakarn2, Pongsak Utaisincharoen3. 1. Department of Oral Microbiology, Faculty of Dentistry, Mahidol University, Bangkok, 10400, Thailand. 2. Department of Microbiology, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand. 3. Department of Microbiology, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand. pongsak.uta@mahidol.ac.th.
Abstract
INTRODUCTION: We aimed to investigate the involvement of surface TLRs and endosomal TLRs in the regulation of SARM expression by TLR2 ligands (Pam2CSK4 and Pam3CSK4). MATERIALS AND METHODS: Mouse macrophage cell line (RAW264.7) was treated with either Pam2CSK4 or Pam3CSK4 (TLR2 ligands) at a concentration of 100 ng/ml. At indicated time points, the treated cells were lysed. The gene and protein expression of SARM were determined by RT-PCR and immunoblotting, respectively. For silencing of TLR9 function, the cells were transfected with TLR9 siRNAs before stimulation by these two TLR2 ligands RESULTS: The SARM expression was upregulated at both transcriptional and translational levels in time-dependent manner during activation of Pam2CSK4 and Pam3CSK4 in mouse macrophages. Blocking of ligand internalization by cytochalasin D showed interference effect with SARM expression. Moreover, our results also demonstrated that endosomal acidification and TLR9 were required for SARM expression suggesting the essential role of endosomal compartment acidification and TLR9 in regulating SARM expression. CONCLUSION: Our findings suggested the collaboration of TLR2-TLR9 at least in the regulation of SARM expression. However, the underlying mechanism that participated in these two TLRs cooperation is underinvestigated.
INTRODUCTION: We aimed to investigate the involvement of surface TLRs and endosomal TLRs in the regulation of SARM expression by TLR2 ligands (Pam2CSK4 and Pam3CSK4). MATERIALS AND METHODS:Mouse macrophage cell line (RAW264.7) was treated with either Pam2CSK4 or Pam3CSK4 (TLR2 ligands) at a concentration of 100 ng/ml. At indicated time points, the treated cells were lysed. The gene and protein expression of SARM were determined by RT-PCR and immunoblotting, respectively. For silencing of TLR9 function, the cells were transfected with TLR9 siRNAs before stimulation by these two TLR2 ligands RESULTS: The SARM expression was upregulated at both transcriptional and translational levels in time-dependent manner during activation of Pam2CSK4 and Pam3CSK4 in mouse macrophages. Blocking of ligand internalization by cytochalasin D showed interference effect with SARM expression. Moreover, our results also demonstrated that endosomal acidification and TLR9 were required for SARM expression suggesting the essential role of endosomal compartment acidification and TLR9 in regulating SARM expression. CONCLUSION: Our findings suggested the collaboration of TLR2-TLR9 at least in the regulation of SARM expression. However, the underlying mechanism that participated in these two TLRs cooperation is underinvestigated.
Authors: Michael Carty; Rory Goodbody; Martina Schröder; Julianne Stack; Paul N Moynagh; Andrew G Bowie Journal: Nat Immunol Date: 2006-09-10 Impact factor: 25.606
Authors: Julianne Stack; Sarah L Doyle; Dympna J Connolly; Line S Reinert; Kate M O'Keeffe; Rachel M McLoughlin; Søren R Paludan; Andrew G Bowie Journal: J Immunol Date: 2014-11-10 Impact factor: 5.422