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Inhibition of ERES cargo loading abrogates ERES recruitment to the inclusion.

By Mary S. Dickinson (6551732), Lindsey N. Anderson (506469), Bobbie-Jo M. Webb-Robertson (46423), Joshua R. Hansen (4180042), Richard D. Smith (39311), Aaron T. Wright (125925) and Kevin Hybiske (129225)

Abstract

<p>(A) Treatment of <i>C. trachomatis</i> infected cells with FLI-06 disrupted the recruitment of COPII coat protein Sec31A to inclusion membranes. Immunofluorescence microscopy of cells showing cellular distribution of Sec31A (anti-Sec31A, first column, green in merges), inclusion membrane protein IncA (anti-IncA, second column, magenta in merges), and DNA (DAPI, third column, blue in merges). Representative deconvolved merged z-series images of uninfected cells are shown in upper panels, and cells infected with <i>C. trachomatis</i> L2 at 24 hpi are shown in lower panels. 10 μM FLI-06 treatment for 4 h, from 20–24 hpi, resulted in Sec31A distribution similar to that of uninfected cells, and away from inclusion membranes. Scale bars = 16 μm. (B) Quantification of Sec31 in cells infected and treated with FLI-06 as described in A. Sec31 punctae that were touching or overlapping with IncA in untreated or FLI-06 treated cells were counted using Volocity to assess the number of overlapping spots per inclusion. At least 20 inclusions were analyzed per condition, for two independent experiments. Each dot represents an inclusion, lines represent mean (SD). Significance determined by unpaired t-test with Welch’s correction, ****, p < 0.0001.</p

Topics: Biochemistry, Medicine, Microbiology, Genetics, Developmental Biology, Cancer, Infectious Diseases, Biological Sciences not elsewhere classified, enzyme APEX 2, Chlamydia trachomatis inclusion membrane, mass spectrometry analysis, endoplasmic reticulum exit sites Chlamydia trachomatis, 20 nm radius, inclusion membrane, ER, epithelial cell infection, 8 hours post infection, ERES, host epithelial cell, inclusion membrane proteome changes, III, endoplasmic reticulum exit sites, APEX 2 system, inclusion membrane proteins, APEX 2
Year: 2019
DOI identifier: 10.1371/journal.ppat.1007698.g006
OAI identifier: oai:figshare.com:article/7948862
Provided by: FigShare
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