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. Author manuscript; available in PMC: 2018 Jan 1.
Published in final edited form as: Cell Microbiol. 2016 Jul 15;19(1):10.1111/cmi.12637. doi: 10.1111/cmi.12637

Figure 4. ORP1L simultaneously co-localizes with the PV and ER.

Figure 4

(A) Live cell confocal microscopy images of ORP1L-GFP on the C. burnetii PV. HeLa cells were transfected with wild type ORP1L-GFP or an FFAT mutant, ORP1L(D478A)-GFP. Three days after infection with mCherry-expressing C. burnetii, the PVs were identified by phase microscopy and fluorescence imaged by live cell spinning disk confocal microscopy. When the top surface of the PV is examined, wild type ORP1L exhibits a striated pattern. This pattern is disrupted by the FFAT D478A mutation, which prevents binding to the ER protein VAP. (B) Live cell fluorescence microscopy images of C. burnetii-infected HeLa cells expressing ER-localized red fluorescent protein (KDEL-RFP) and ORP1L-GFP. The maximum Z projection (Z projection) shows the flattened confocal stack through the entire cell, while the PV surface is a confocal slice of the top surface of the PV (arrow). As shown in the magnification of the boxed PV, ORP1L-GFP on the PV co-localized with the host ER. Red = ER, Green = ORP1L-GFP. Scale bars = 10 µm.