Confocal microscopy images of selected cell markers in infected DH82 cells.

Abstract

<p>When the infectivity reached 95%, the infected cells were fixed and permeabilized. The slides were incubated with the primary antibodies specific to the selected makers (anti-CD71, anti-EEA1, anti-Rab5, anti-LAMP II, anti-Rab7, anti-VATPase, anti-MAP LC3, or anti-BECN antibodies), and then incubated with corresponding secondary conjugated antibodies. In all images, the blue indicated DAPI-stained nuclei (large blue body) or <i>E. chaffeensis</i> morulae; the red indicated host cell proteins labeled with antibodies to CD71 (A), EEA1 (B), Rab5 (C), LAMP II (D), Rab 7 (E), cathepsin D (F), VATPase (G), MAP LC3 (H), and BECN (I). Uninfected DH82 cells incubated with primary antibodies and corresponding secondary antibodies (J) and infected DH82 cells incubated with secondary conjugated antibodies alone (K) served as negative controls. In each panel, whether the DAPI staining was co-stained with host cell proteins determined the co-localization of <i>E. chaffeensis</i> and the host cell proteins. These results were confirmed in three independent experiments.</p