Figure 12.

Endogenous and GFP-tagged OPTN can be recruited to peroxisomes under basal growth conditions. DD-DAO Flp-In T-REx 293 cells were co-transfected (lower panels) or not co-transfected (upper panels) with plasmids encoding OPTN-GFP and mCherry-PTS1. After culturing the cells for two days in rMEMα, they were processed for live-cell imaging (lower panels) or fixed and processed for immunofluorescence microscopy using a mouse antiserum against PEX14 and a rabbit antiserum against OPTN, in combination with goat anti-mouse and anti-rabbit secondary antibodies conjugated to Alexa Fluor 488 and Texas Red, respectively (upper panels). The white arrowheads point to sites of colocalization between OPTN(-GFP) and the peroxisomal marker protein PEX14 or mCherry-PTS1. Scale bar, 10 µm.