Journal: Nature Biotechnology

Article Title: A highly photostable and bright green fluorescent protein

doi: 10.1038/s41587-022-01278-2

Figure Lengend Snippet: a , Labeling microtubule plus-end with tdStayGold. left , Domain structure of EB3 = tdStayGold. EB3: Microtubule (MT)-associated end-binding protein 3. right , COS-7 cells expressing EB3 = tdStayGold were continuously imaged for 30 min (see Supplementary Video ). Exposure time: 0.5 s. The first and last fluorescence images are shown. Comparison of the two images reveals that there was no substantial photobleaching during the 30-min observation. Scale bar, 20 μm. The EB3 dynamics shown are representative of n = 3 similar observations. b , Labeling postsynaptic density (PSD) with tdoxStayGold. top , Domain structure of PSD-95=tdoxStayGold. bottom , Cultured neurons prepared from embryonic rat hippocampus were cotransfected with the cDNAs of PSD-95=tdoxStayGold and mCherry at DIV (days in vitro) 5. Transfected neurons were imaged at DIV 25 in DMEM/F12 (1:1) containing 2% (vol/vol) FBS, N2-supplement (1×), and B-27 (1.5×) using the SpinSR10. Volumetric imaging was performed in z steps of 0.25 µm. The imaged volume comprised 30 z slices. Areas enclosed by white boxes are enlarged on the right side. Scale bars, 1 µm. Similar images were obtained from 6 other imaging experiments performed at DIV 21–25 on 4 independent neuronal samples. a , b , =: Coupler linker, a triple repeat of the amino acid linker Gly-Gly-Gly-Gly-Ser.

Article Snippet: Then, fresh growth medium DMEM (041-29775, Fuji Film) (10% FBS, penicillin–streptomycin) was substituted.

Techniques: Labeling, Binding Assay, Expressing, Fluorescence, Comparison, Cell Culture, In Vitro, Transfection, Imaging, Ubiquitin Proteomics