Sub-cellular co-localization of gTLR9A, gTLR9B, and gMyD88 in the endosome.

<p>GK cells were co-transfected with gTLR9A-Flag and YFP-gMyD88 (<i>A</i>–<i>D</i>) or gTLR9B-HA with YFP-gMyD88 (<i>E</i>–<i>H</i>). Transfected cells were incubated with the endosome marker transferrin conjugated to Alexa Fluor 633 along with CpG ODN for 30 min. Images were then acquired on an inverted confocal microscope. The merged images (<i>D</i> & <i>H</i>) demonstrate that gTLR9A and gTLR9B can translocate to the endosome and co-localize with gMyD88 upon stimulation with CpG ODN. The sub-cellular co-localization of CpG ODN and the receptor-adaptor complex was assayed in GK cells transfected with EYFP-MyD88 and treated with Texas-red-conjugated ODN. Both ODN (<i>I</i>) and gMyD88 (<i>J</i>) formed granular aggregates and co-localized with each other (<i>K</i>). The fluorescent granules exhibited structural features of organelles under differential interference contrast (DIC) microscopy (<i>L</i>). Scale bar: 10 μm. Results present the observation of 100 transfected cells or more from one representative experiment out of two.