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Interference with FAK functions results in decreased uptake of <i>N. meningitidis.</i>

By Heiko Slanina (154782), Sabrina Hebling (154785), Christoph R. Hauck (154786) and Alexandra Schubert-Unkmeir (154787)

Abstract

<p>(A) 293T cells were transfected with a control plasmid (pcDNA) and a plasmid encoding wildtype FAK (HA-FAK WT) or a plasmid encoding a kinase inactive mutant (HA-FAK K454M) or a mutant that was not capable of autophosphorylation (HA-FAK Y397F). Transfected cell were infected with invasive strain MC58 <i>siaD</i> and intracellular bacteria were estimated 4 h post-infection by gentamicin protection assays. The graph represents mean values ± S.D. of three independent experiments done in duplicate. * <i>P</i><0.05, relative to cells transfected with the control plasmid, <sup># </sup><i>P</i> ≤ 0.05, relative to cells transfected FAK wildtype were considered significant. In parallel, Western blotting of WCL extracts with anti-HA-tag antibody demonstrates expression of HA-tagged FAK constructs. (B) 293T cells were transfected with HA-FAK WT or a plasmid encoding the FAK related non-kinase (HA-FRNK) and infected with MC58 <i>siaD</i> as described above. The graph represents mean values ± S.D. of three independent experiments done in duplicate. * <i>P</i><0.05. WCL were analyzed by Western blotting using an anti-HA-tag antibody and demonstrated overexpression of FRNK in transfected cells.</p

Topics: Microbiology, Cell Biology, Molecular Biology, Infectious Diseases, fak, functions, uptake
Year: 2012
DOI identifier: 10.1371/journal.pone.0039613.g003
OAI identifier: oai:figshare.com:article/286339
Provided by: FigShare
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