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. 2012 Nov 16;1(4):1121–1132. doi: 10.3390/cells1041121

Figure 1.

Figure 1

Cell architecture and organization of subcellular compartments depend on the experimental model—human submandibular glands (HSG) cells were cultured either on glass (2D, upper panels) or on 5 mg/mL matrigel (3D, center panels), as described in Material and Methods. (A) Phase contrast images of HSG cells grown in 2D (upper panel) or in 3D (center panel). In 3D cells form acinar-like structures that resemble the acini in intact SGs. In the lower panel an intact rat submandibular gland was imaged by two-photon microscopy (Excitation 740 nm) as previously described [17]. An acinus is highlighted (red broken line). (B) HSG cells grown in 2D or 3D and intact salivary glands were labeled with Hoechst (blue) and Texas-Red Phallodin (red) to highlight nuclei and actin respectively. Z-scans were acquired by confocal microsopy (excitation 405 nm and 562 nm) and maximal projections of the stacks are shown. In 2D, actin is mainly localized in stress fibers (upper panels, arrow), whereas in 3D is localized at the plasma membrane (center panel, arrow). In intact SGs, actin is localized primarily at the apical plasma membrane and decorates the acinar canaliculi (lower panel, arrows). Bars, 10 μm. (C) HSG cells and intact SGs were labeled with antibodies against VAMP8 (red) and TfnR (green). HSG cells were also labeled with Hoechst (blue), whereas SGs were labeled with phalloidin (Cyan). Note that in SGs VAMP8 is localized close to the apical plasma membrane and does not colocalize with TfnR (lower panel). Bars, 10 μm.