Skip to main content
. 2014 Jul 3;10(7):e1004222. doi: 10.1371/journal.ppat.1004222

Figure 5. The effector Lpg1101 requires both its C-terminal LEPR and MIM regions for vacuolar localization.

Figure 5

(A) Confocal micrograph of HEK293 cells stably expressing both FcγRII and 3XFLAG-tagged Lpg1101 and infected with wild type Lp01. The localization of 3xFLAG-tagged Lpg1101 was detected by indirect immunofluorescence using indirect the affinity purified anti-Lpg1101 antibody (1∶100) and secondary Alexa Fluor 488 (green). L. pneumophila were detected due to constitutive expression of mCherry (red). Lpg1101 was detected at both at the cell periphery and surrounding Legionella. (B) Quantification of the localization of EYFP-tagged Lpg1101 and LEPR and MIM mutant variants with intracellular Lp01 at 30, 60 and 120 min post-infection. Amino acid substitutions are shown in red. (C) Fluorescent micrographs of the localization of EYFP-Lpg1101 (EYFP1101) and the LEPR (D243E/K246R) and MIM (L288A/V292A/T293A/I295A) variants in cells infected with mCherry expressing L. pneumophila strain Lp01 (red). Hoechst 33343 was used to stain DNA (blue).