Subcellular Localization of RTT109-EGFP Fusion Proteins

To investigate the subcellular localization of RTT109-EGFP fusion proteins, the strains were cultivated on glass slides along with strains expressing H3-mRFP fusion proteins, as previously described (Schmidt et al. 2020 (link)). Heterokaryotic strains can therefore express both fluorescence-labeled proteins (Engh et al. 2007 (link)). Subsequently, light and fluorescence microscopy were performed using an AxioImager microscope (Zeiss), equipped with a Photometrix Cool SnapHQ camera (Roper Scientific). EGFP fluorescence was detected using a Chroma filter set 41017 (HQ470/40, HQ525/50, Q495lp), while mRFP fluorescence was detected using set 49008 (EG560/40x, ET630/75m, T585lp). The acquired images were further processed using MetaMorph software (Molecular Devices).