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Text S1 - A Role for <i>LHC1</i> in Higher Order Structure and Complement Binding of the <i>Cryptococcus neoformans</i> Capsule

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Supporting information file containing Table S1 showing sequences of DMSO-solubilized proteins, Table S2 showing glycosyl composition analysis (mole %), Figure S1 showing capsule diameter is dependent on LHC1, Figure S2 showing induction of Lhc1-mCherry in ASN and RPMI media, Figure S3 showing quantitative RT-PCR of LHC1 under indicated conditions, Figure S4 showing aggregate TMRD diffusion measurements, Figure S5 showing western blot of cryptococcal extract using anti-Lhc1 antigen-purified antibody, Figure S6 showing deletion of LHC1 results in increased iC3b binding from human serum. And Supplemental Materials and Methods containing fungal strains, plasmids and media, disruption and complementation of LHC1 in C. neoformans, Lhc1-mCherry fusion protein, quantitative RT-PCR experiments, western blot analysis, analysis of lactonohydrolase activity, preparation of a recombinant fragment and full-length lactonohydrolase and generation of anti-Lhc1 antibody, Immunolocalization studies, biophysical analysis of DMSO-extracted capsular polysaccharide, Cryo-Scanning Electron Microscopy (Cryo-SEM). Immunolocalization studies, phagocytosis assay, fungal killing assay by human monocytes, fungal killing by J774A.1 cells, mouse virulence model with the addition of cobra venom factor, detection of C3 and iC3b binding by flow cytometry, and Supplemental References. (DOCX)

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2014-05-01
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