Goat anti rabbit and goat anti mouse hrp conjugated secondary antibodies

Rabbit anti-human Notch1 (NICD); mouse anti-human Numb, E-cadherin, N-cadherin, and Slug; and goat anti-human Snail primary antibodies were obtained from Abcam (Beverly, MA, USA). Goat anti-rabbit and goat anti-mouse HRP-conjugated secondary antibodies and Diaminobenzidine staining kits were obtained from ZSGB-BIO (Beijing, China).
Normal endometria were obtained from 45 women of reproductive age undergoing bilateral tubal ligation (proliferative phase: n = 15; secretory phase: n = 15; postmenopausal phase: n = 15). Adenomyotic lesions were obtained from 30 patients with adenomyosis undergoing hysterectomy or subtotal hysterectomy (proliferative phase: n = 15; secretory phase: n = 15) (Table 1). Normal endometria and adenomyosis tissues were collected during the operation.The diagnosis of adenomyosis was confirmed by histological examination. No patients received any hormonal therapy in three months prior to their surgery. Informed consent was obtained from all participants prior to the biopsy procedure, and the use of human tissues was approved by the institutional review board of Shandong Provincial Hospital Affiliated to Shandong University.Table 1

Detailed information of patients

Proliferative phase		Secretory phase
Ages	Dysmenorrhea	Ages	Dysmenorrhea
39	+	47	+
44	+	41	+
47	+	50	+
44	+	45	+
45	+	42	+
43	+	42	+
32	+	47	+
47	+	37	+
37	+	40	+
43	+	48	+
47	+	43	+
44	-	52	+
49	+	45	+
39	+	48	+
50	+	51	+

CollagenaseΙA, trypsin, Melatonin and Matrigel were obtained from Sigma-Aldrich (St. Louis, MO, USA). Penicillin, DMEM/F12 (1:1) media were obtained from HyClone (Logan, Utah, USA). Charcoal-stripped fetal bovine serum (FBS) was obtained from GIBCO (Invitrogen, NY, USA). Rabbit anti-human E-Cadherin, N-Cadherin and Vimentin primary antibodies were obtained from Abcam (Cambridge, MA, USA). Rabbit anti-human Notch, Numb, Slug and Snail primary antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA) for western blot and obtained from Abcam (Cambridge, MA, USA) for immunohistochemistry. Mouse anti-human β-Actin primary antibody and Goat anti-rabbit and Goat anti-mouse HRP-conjugated secondary antibodies were obtained from ZSGB-BIO (Beijing, China). Mammalian Cell Protein Extraction Kit was purchased from Beyotime (Shanghai, China). ECL Plus Western Blotting Detection System was obtained from Millipore Corporation (Billerica, MA, USA).