Figure 7

Silencing of aggrephagy-associated proteins affects the autophagosomal degradation of chlamydial structures in infected DCs. (A) Infected DCs (48 hpi) were costained for Parkin, p62, or ubiquitin (red), chlamydial structures (green) and DNA (blue) (left panel). Enlarged photographs (I, II and III, right panel) show colocalisation between aggrephagy-associated proteins and chlamydial structures. (B) The fluorescence intensity along a cellular cross section of interest was measured. Obtained profiles were overlaid and coloured. (C) DCs were siRNA-silenced for Parkin, HSP25/27 and HDAC6 and then infected (48 hpi) with chlamydia. siRNA silencing of Parkin, HSP25/27 and HDAC6 was demonstrated by Western blot (upper panels). Asterisk indicates an unspecific cross-reacting band. Autophagy induction in non-infected and infected (48 hpi) DCs (silenced for the different proteins) was analysed by CytoID staining (CAT). Fluorescence profiles were overlaid to directly compare changes in autophagy (lower panels). Data from three independent experiments (non-infected and 48 hpi) are summarised as bar graphs with arbitrary units (D). The value obtained for non-infected cells was set to 1. Statistical analysis in (D) was performed as described in Methods (*p < 0.05; **p < 0.01; ***p < 0.001 versus controls; n = 3).