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. Author manuscript; available in PMC: 2016 May 1.
Published in final edited form as: Arch Oral Biol. 2015 Feb 25;60(5):715–723. doi: 10.1016/j.archoralbio.2015.01.010

Figure 2.

Figure 2

Immunofluorescence localization of Gαs in mouse parotid gland. Scale bars: main panels, 50 μm; insets, 20 μm. (For color reproduction on the Web and in print.)

A. Unstimulated control. In the secretory acinar cells (AC), Gαs is localized to the basolateral membranes; minimal cytoplasmic labeling is observed. Cells of the striated (SD) and intercalated (ID) ducts show strong cytoplasmic fluorescence. Smooth muscle cells surrounding small blood vessels (BV) also are labeled.

B. Unstimulated control incubated with non-immune IgG. Weak non-specific fluorescence is seen in the acini and ducts. Lysosomes exhibit orange autofluorescence.

C. Isoproterenol, 15 min. Gαs fluorescence is seen along the cell membranes and present in the cytoplasm of the acinar cells (AC). No fluorescence is present in the nuclei. Apical fluorescence in the striated ducts (SD) appears to be increased.

D. Isoproterenol, 30 min. Most of the fluorescence in acinar cells (AC) is cytoplasmic, and membrane-associated fluorescence is further reduced. Apical fluorescence is still present in the striated ducts (SD).

E. Isoproterenol, 1 hr. Acinar cell (AC) cytoplasm remains fluorescent, however, the apical labeling of striated ducts (SD) appears reduced (inset). A similar distribution of fluorescence was seen at 2 hr and 4 hr after isoproterenol administration.

F. Propranolol/Isoproterenol, 1 hr. Administration of the β-receptor antagonist propranolol 30 min prior to isoproterenol reduces redistribution of Gαs. Cytoplasmic fluorescence of acinar cells (AC) is reduced, and some Gαs remains associated with the cell membranes. Striated duct (SD).

G. Isoproterenol, 6 hr. The acinar cells (AC) exhibit decreased cytoplasmic fluorescence and increased membrane-associated fluorescence. Apical fluorescence of striated duct (SD) cells is reduced.

H. Isoproterenol, 8 hr. The distribution of fluorescence is similar to that of the unstimulated control gland. Most acinar cells (AC) show basolateral membrane-associated fluorescence, and minimal cytoplasmic fluorescence. Intense cytoplasmic fluorescence is seen in striated ducts (SD);.apical fluorescence is diminished. Blood vessel (BV).