Ab17204

Total protein was collected from normal endometrial tissues, eutopic and ectopic endometrial tissues with EMS, or cells with RIPA Lysis and Extraction Buffer (Thermo Scientific). Approximately 20 μg of total protein was loaded on 12% sodium dodecyl sulfate-polyacrylamide gels (SDS-PAGE) and transferred to a 0.2 μm PVDF membranes (Roche, Basel, Switzerland). The blots were incubated overnight at 4°C with primary antibodies against FBLN1 (1 : 1000, ab17204, Abcam), EFEMP1 (1 : 1000, ab256457, Abcam), and β-actin: (1 : 2000, ab8227, Abcam) and were then incubated with a goat anti-rabbit HRP-conjugated secondary antibody. The protein signals were visualized by an ECL kit (Solarbio).

Cells were fixed with 4% formaldehyde, permeabilized with 0.2% Triton X-100 in PBS, and blocked with 2% BSA and 0.1% Triton X-100 in PBS. Cells were incubated overnight with anti-XAF1 (ab17204, Abcam) or anti-IRF-1 (SC-514934) antibody at 4 °C, and stained with secondary antibodies. After mounting the coverslips, fluorescent imaging was obtained with a confocal laser scanning microscopy (LSM700, Carl Zeiss MicroImaging, Inc).

Rabbit antibodies against XAF1 (ab17204) were purchased from Abcam (Cambridge, England). Rabbit antibodies against β-actin (4970) and HRP-conjugated anti-rabbit (7074) were purchased from Cell Signaling Technology (Danvers, USA). Enhanced chemiluminescence (ECL) HRP substrate was supplied by Thermo (Fremont, USA). The plasmid of pcDNA3.1 and Lipofectamine 2000 were purchased from Invitrogen (Carlsbad, USA). The plasmids of pGL3-promoter and pRL-SV40 luciferase reporter were purchased from Promega (Madison, WI, USA). The shRNA expression plasmids of pGPU6-GFP-Neo vector were from GenePharma (Shanghai, China). QIAprep spin miniprep kit was obtained from Biomiga (San Diego, CA, USA). An annexin V-APC/PI kit was purchased from Bender MedSystems (Vienna, Austria). Cell Counting Kit-8 (CCK-8) was form MedChemExpress (Monmouth Junction, NJ, USA).