Rabbit anti cd133

The human prolactinoma specimens (n=14) and rat xenograft tumors (n=9) were fixed, paraffin-embedded, and cut into 4-μm-thick sections. The sections were deparaffinized, rehydrated, and rinsed with PBS, followed by antigen retrieval in citrate buffer (pH 6.0) for 15 min at 95–100°C. Subsequently, non-specific antigens were blocked in 5% donkey serum for 60 min at 25°C. Then sections were incubated with primary antibodies (Rabbit anti-CD133, 1: 100, MyBioSource, USA; Goat anti-Nestin, 1: 50, Santa Cruz, USA; Rabbit anti-Oct4, 1: 100, Proteintech, USA; Rabbit anti-Sox2, 1: 100, Proteintech, USA; Mouse anti-D2DR, 1: 50, Santa Cruz, USA) overnight at 4°C. After being rinsed with PBS, sections were incubated with secondary antibodies (FITC or PE conjugated Donkey anti-Rabbit, anti-Mouse or anti-Goat IgG, 1: 200, Santa Cruz, USA) for 60 min at 25°C. Finally, sections were counterstained with DAPI (Sigma, USA) and mounted with Antifade Mounting Medium (Beyotime,China) before being examined by fluorescence microscopy (Olympus, Tokyo, Japan).

After cabergoline treatment, MMQ cells were collected and washed with PBS. Then, MMQ cells were resuspended in 100 μL staining buffer (LiankeBio, China) and incubated with Rabbit anti-CD133 (1: 50, MyBioSource, USA) for 30 min at 4°C. Negative control was incubated with equivalent PBS instead. After washing with PBS, cells were incubated with FITC-conjugated Donkey anti-Rabbit IgG (1: 50, Santa Cruz, USA) for 30 min at 4°C. Subsequently, cells were washed and resuspended in PBS. Then, the proportion of CD133-expressing cells in all groups was analyzed by a FACS Calibur Flow Cytometer (BD Biosciences). All experiments were repeated 3 times.