Fig. 3.

Depletion of sortilin does not impact upon delivery of ceramide to the inclusion. (A) The protein levels of sortilin in stable cell lines expressing non-silencing shRNA or shRNA targeting sortilin were analysed by western immunoblotting. (B) Sortilin knockdown cells were infected with CTL2 (MOI 0.5). At 23 hpi, cells were treated with BODIPY-FL-ceramide and delivery of ceramide followed for 4 h via live cell microscopy. Arrows highlight chlamydial inclusion. Images were captured using a Nikon Ti-E deconvolution microscope with a 40× objective. (C) Line graph represent the shift in mean fluorescent intensity per inclusion analysed by ImageJ. At least 9 inclusions were monitored. Error bar denote mean±s.e.m., N=2. Scale bar 20 µm.