Immunohistochemistry staining

Immunohistochemistry (IHC) staining was carried out by using the 2-step PV-9000 kit (ZSGB-BIO, China) as described previously [12]. To be briefly, paraffin-embedded tissues were cut into 8-μm serial sections and dried at 70°C for 2 hours. The sections were then washed 3 times with PBS (phosphate buffered saline) and deparaffinized with xylene and rehydrated with alcohol. The antigen retrieval was carried out in a microwave oven using sodium citrate buffer. Specimens were then incubated with 10% serum to block nonspecific binding after washing 3 times with PBS. The incubation with a rabbit anti-human GPR110 antibody (1: 500 dilution; ab150547; Abcam) was performed at 4°C overnight. Then the sections were washed and incubated with the Polymer Helper and using poly peroxidase-anti-rabbit IgG was used for detection and sections mounted with a neutral resin. Negative control staining was carried out with PBS.