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. 2010 Apr 27;20(8-6):703–709. doi: 10.1016/j.cub.2010.02.049

Figure 3.

Figure 3

Intracellular Localization of SpTPCs

(A) Colocalization of mCherry.SpTPC with acidic organelles (LysoTracker Green) in live HEK293 cells expressing mCherry.SpTPCs. Scale bar corresponds to 10 μm.

(B) Anti-SpTPC3 immunofluorescence of S. purpuratus eggs with or without peptide block for assessment of immunostaining specificity. Scale bar corresponds to 10 μm, or 2 μm for higher magnification panel.

(C) Localization of SpTPCs.mCherry expressed in oocytes of starfish Asterina miniata and detected by mCherry fluorescence or by immunofluorescence with corresponding anti-SpTPC antibodies. Scale bar corresponds to 20 μm. Noninjected controls were not labeled with anti-SpTPC antibodies (data not shown). In oocytes expressing SpTPC3.mCherry, the germinal vesicle is out of the plane of focus. mCherry.SpTPCs showed similar cortical localization (data not shown). Oocytes expressing mCherry alone showed a signal across the entire cell (data not shown). Of note is the presence of an SpTPC1 signal in the nucleus. Indeed, SpTPC1 has a putative nuclear localization signal (amino acids 327–334). Its physiological significance may represent an important avenue for further analysis of TPC function, especially in light of reports describing NAADP signaling in nucleus [33, 34]. See also Figure S3.