Fig. 5. Localization of TPC2 with respect to the nucleus and peri-nuclear region of SMCs.

Single optical sections (taken at the widest part of the nucleus) to show the localization of TPC2 and other proteins in the nuclear region of SMCs. Cells were dual-immunolabeled with: (A,B) myosin heavy chain and TPC2 antibodies; (C,D) myosin heavy chain and LAMP1 antibodies; (E,F) RyR and IP₃R type III antibodies, (G) myosin heavy chain and IP₃R type I antibodies or (H) myosin heavy chain and IP₃R type II antibodies, and then counterstained with DAPI to label the nucleus. In panels A-D, G and H the myosin heavy chain antibody was simply used to identify SMCs in the mixed cell culture (and is therefore not shown). In the case of TPC2, LAMP1, IP₃R type I and IP₃R type II, the localization pattern of these proteins was shown alone (A-D,G,H, panel a) and when superimposed on the image of the nucleus (A-D,G,H, panel c). In the case of RyR and IP₃R, the localization pattern of each protein was shown alone (E,F panels a and b, respectively), and when superimposed together and with the image of the nucleus (E,F panel d). Scale bars, 5 μm. (Ad-Dd,Ee,Fe,Gd,Hd) Line-scan analyses. The 0 and 18 in panel Ac indicate the start and end point of the line-scan, which makes up the x-axis of panel Ad.