Online Supplementary Data
CMRF-35-like molecule 1 (CLM-1) regulates eosinophil homeostasis by suppressing cellular chemotaxis
Itay Moshkovits 1, Dana Shik 1, Michal Itan 1,Danielle Karo-Atar 1, Biana Bernshtein 2, Alon Y Hershko 3, Menno van Lookeren Campagne 4, Ariel Munitz 1,*
1Department of Clinical Microbiology andImmunology, The Sackler School of Medicine, Tel-Aviv University, Ramat Aviv 69978, Israel.
2Department of Immunology, Weizmann Institute of Science, Rehovot 76100,
Israel
3Laboratory of Allergy and Clinical Immunology, Department of Medicine, The Herbert Center of Mast Cell Disorders, Meir Medical Center, Kfar Saba, Israel.
4Department of Immunology, Genentech Inc., South San Francisco, CA 94080
Running Title: CLM-1 regulates eosinophil migration
Scientific category: Phagocytes
Supplemental figure count: 3
*Corresponding author: Ariel Munitz, PhD, Department of Clinical Microbiology andImmunology, The Sackler School of Medicine, Tel-Aviv University, Ramat Aviv 69978, Israel. Tel. (Office): +972-3-640-7636, Fax: +972-3-640-9160, e-mail: .
Grant support:This study was supported by research funding toA.Y.H. (Morasha Program,Israel Science Foundation, grant no. 1084/10); to I.M the Joan and Jaime Constantiner Institute for Molecular Genetics; to A.M. by the FP7 Marie-Curie Reintegration grant (grant no. 256311), theUS-Israel Binational Science Foundation (grant nos. 2009222 and 2011244), the Israel Science Foundation (grant no. 955/11), the Israel Cancer Research Foundation Research Career Development Award, the Dream Ideas Djerassi-Elias Institute of Oncology Research Fund, and internal Tel-Aviv University funds.
Figure legends
Figure S1. Expression of CLM-1 in alveolar macrophages. Bone marrow (BM), spleen and whole lung samples were obtained from naïve wild type (WT) mice. Lung samples were subjected to enzymatic digestion to generate single cell suspensions. Thereafter, the cells were stained and CD45+ cells gated (A, “R1”). Alveolar macrophages or eosinophils were defined as CD45+/Gr-1int/CD11c-/Siglec-F+/CD11b+- “R2” and CD45+/Gr-1-/CD11c+/Siglec-F+/CD11b-- “R3”, respectively. Representative histogram plots of CLM-1 expression on the surface of eosinophils (B) alveolar macrophages (C), BM and splenic macrophages (D and E, resepectively) is shown.
Figure S2. Expression of CCR3 and eotaxins in WT and Clm1-/- eosinophils and tissues, respectively.CCR3 expression on the cell surface of eosinophils from the indicated anatomical locations was assessed by flow cytometry (A). Eotaxin-1 (B) and eotaxin-2 (C) levels in the indicated organs was assessed by ELISA; PL, peritoneal lavage; n = 3; ns, non significant; *P < 0.05.
Figure S3. Expression of CD11b in wild-type and Clm1-/-eosinophils following eotaxin stimulation.The expression of CD11b on the cell surface of WT and Clm1-/- eosinophils was assessed following eotaxin stimulation; n = 3; NA, non-activated, ns, non-significant.
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