<i>Brucella</i> impairs host cell secretion during infection.

<p>(A) Quantification of SEAP secretion in uninfected (control) and <i>B. abortus</i> 2308-infected (2308) HeLa cells. HeLa cells were transfected with pSEAP-2 for 16 h, then infected with <i>B. abortus</i> 2308. At 24 h pi, SEAP protein production was synchronized as described in the <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003556#s4" target="_blank">Materials and Methods</a> and the external and internal SEAP activity measured at various times post-synchronization. Data show SEAP ratios (external/internal SEAP activity) from uninfected or infected cells. (B–F) VSV-Gts045-GFP trafficking in either control or <i>B. abortus</i> 2308-infected HeLa cells. HeLa cells were either mock-infected or infected with DsRed_m-expressing <i>B. abortus</i> 2308. Twenty-four hours pi, cells were transfected with plasmid encoding VSV-Gts045-GFP and incubated at 40°C for 20 h. After 30 min of cycloheximide treatment to stop VSV-Gts045-GFP production, cells were shifted to 32°C, then fixed and immunostained at the indicated time points and VSV-Gts045-GFP localization was examined. (B) Percentage of VSV-Gts045-GFP in the ER, (C) in the ERGIC, (D) in the Golgi apparatus, (E) in the Golgi to plasma membrane (Golgi-PM) compartments and (F) % of VSV-G at the plasma membrane (PM). Data are expressed as a percentage of where VSV-G is located in the defined compartment, as shown in the representative confocal micrographs and are the means ± SD from three independent experiments.