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. 2014 Oct 17;9(10):e110544. doi: 10.1371/journal.pone.0110544

Figure 2. Intracellular localization of Dictyostelium GFP-Cln3 using epifluorescence microscopy.

Figure 2

(A) AX3 cells overexpressing GFP-Cln3 imaged live in water. Scale bar = 5 µm. (B) AX3 cells overexpressing GFP-Cln3 were fixed in either ultra-cold methanol (for VatM and Rh50 immunostaining) or 4% paraformaldehyde (for p80 immunostaining) and then probed with anti-VatM, anti-Rh50, or anti-p80, followed by the appropriate secondary antibody linked to Alexa 555. Cells were stained with DAPI to reveal nuclei (blue). Images were merged with ImageJ/Fiji. VC, vacuolar-shaped structures; VS, cytoplasmic vesicles; T, tubular-like structures within the cytoplasm; P, punctate distributions within the cytoplasm. Scale bars (B, C) = 2.5 µm.