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A secreted MIF homologue from Trichinella spiralis binds to and interacts with host monocytes 1 1 Introduction 1 2 Materials and methods 2 2.1 Ethics statement 2 2.2 Parasites and experimental infection 2 2.3 RNA extraction and cDNA cloning 2 2.4 Expression and purification of recombinant TsMIF 2 2.5 Generation of anti-TsMIF polyclonal antibody 3 2.6 Western blot analysis 3 2.7 Immunofluorescence assay (IFA) 3 2.8 Fluorescent immunohistochemistry assay 3 2.9 GST pulldown assay 3 2.10 Molecular modeling for TsMIF binding to CD74 or Jab1 3 2.11 Western blot detection of ERK1/2 phosphorylation 3 2.12 BrdU cell proliferation ELISA assay 3 2.13 Statistical analysis 4 3 Results 4 3.1 Production of rTsMIF protein and its antiserum 4 3.2 IFA reveals the differential expression and location of TsMIF in different stages 4 3.3 The secreted TsMIF binds to host monocytes 4 3.4 TsMIF interacts with the host cell surface receptor CD74 5 3.5 TsMIF stimulates the activation of ERK1/2 and proliferation in RAW264.7 cells 7 3.6 TsMIF interacts with human Jab1 7 4 Discussion 8 CRediT authorship contribution statement 9 Declaration of Competing Interest 9 Data availability 9 Acknowledgment 9 Funding 9 S

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UND

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nonfiction