4 6 diamidino 2 phenylindole dapi mounting solution

Methyl alcohol (product no. 62), n-hexane (product no. 4794), ethyl acetate (product no. 2936), and chloroform (product no. 1268) were purchased from Duksan chemicals, South Korea. 1-butanol (cat# 33065) was purchased from Honeywell research chemicals. 5-Fluorouracil (cat# F6627), propidium iodide (PI), and 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Dihydrorhodamine 123 (DHR 123: cat# D23806) was purchased from ThermoFisher scientific. Antibodies [cleaved caspase-3 (cat# sc-22171), cleaved PARP (cat# sc-56196), and β-actin (cat# sc-47778)] were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). 4′,6-diamidino-2-phenylindole (DAPI) mounting solution (Cat# H-1200) was purchased from Vector laboratories, USA. All the chemicals and reagents were used as directed by the manufacturers.

The frozen sections were immunostained with anti-macrophage/monocyte antibody (MOMA-2) After washing with PBS, the sections were reacted with Alexa Fluor 488 (goat anti-mouse Ig) and washed. The tissues were reacted with Cy3-conjugated anti-α-smooth muscle antibody (SMA) for 1h at RT, washed thrice with PBS, and then mounted with 4′,6-diamidino-2-phenylindole (DAPI) mounting solution (Vector Labs, Burlingame, CA, USA).

Four weeks after transplantation, the rats were sacrificed with an overdose of isofulrane and the mandible block was removed. The block was fixed in 4% paraformaldehyde-phosphate buffer for 2 days and decalcified in 10% EDTA at 4 °C for 4 weeks, followed by dehydration, paraffin embedding, and sectioning. The sections were observed microscopically using a BZ800 apparatus (Keyence, Osaka, Japan) after staining with hematoxylin-eosin and azan. For labeled cell observation, the section was mounted with 4′,6-diamidino-2-phenylindole (DAPI) mounting solution (Vector Laboratories Inc., Burlingame, CA, USA).