At 14 days after the transplant of salivary organoids,
the submandibular
glands were extirpated. The sections (4 um thickness) of formalin-fixed,
paraffin-embedded tissues were cut and stained with hematoxylin and
eosin (H&E). For immunofluorescence staining of salivary gland
tissues, paraffin sections were dewaxed in xylene for 30 min, after
which antigen retrieval was performed by boiling in 10 mM sodium citrate
for 2 min. The tissue slices were then incubated with a primary antibody
(anti-AQP5 antibody; Alomone Lab, anti-CK18 antibody: Santa Cruz Biotechnology)
overnight at 4 °C, and next they were incubated with a secondary
antibody, an Alexa-conjugated anti-IgG antibody, for 1 h at room temperature
with 4′,6-diamidino-2-phenylindole (DAPI). After mounting,
the cells were viewed under a confocal laser scanning microscope (LSM
780, Carl Zeiss).
the submandibular
glands were extirpated. The sections (4 um thickness) of formalin-fixed,
paraffin-embedded tissues were cut and stained with hematoxylin and
eosin (H&E). For immunofluorescence staining of salivary gland
tissues, paraffin sections were dewaxed in xylene for 30 min, after
which antigen retrieval was performed by boiling in 10 mM sodium citrate
for 2 min. The tissue slices were then incubated with a primary antibody
(anti-AQP5 antibody; Alomone Lab, anti-CK18 antibody: Santa Cruz Biotechnology)
overnight at 4 °C, and next they were incubated with a secondary
antibody, an Alexa-conjugated anti-IgG antibody, for 1 h at room temperature
with 4′,6-diamidino-2-phenylindole (DAPI). After mounting,
the cells were viewed under a confocal laser scanning microscope (LSM
780, Carl Zeiss).