Fig. 4.

Live-imaging of non-fixed cells over-expressing SCAPER, presenting tubular structure: a, b Overexpression of the N’ mEGFP tagged SCAPER wild-type or mutant p.(L936*) truncated protein in NIH/3T3 cells (a) and SH-5YSY cells (b): diffuse and cilia-like or basal body-like organelle localization is evident, as well as a tubular structure for the mutant construct in both cell lines. c, d Co-localization studies showing that the tubular structure expressing SCAPER is indeed a tubulin structure: overexpression of the N′ mCherry-tagged SCAPER mutant p.(L936*) truncated protein in NIH/3T3 cells (c) and SH-5YSY cells (d), co-expressed and co-localized with eGFP tagged alpha-tubulin in live cells. Note: cells were visualized using ZEISS LSM880 microscope with a 40×/1.2 objective, in FastAiryScan mode, in 2 or 3 separate tracks, as described in the methods section. The scale bar equals 10 µm. Cells were imaged 12 h post transfection