Anti cd7

After dewaxing and hybration, the sectioned samples were blocked with 10% FBS. The sections were incubated with primary antibodies at 4°C overnight. The antibodies used were anti-CD7, anti-CD8, anti-CD14, anti-CD15, anti-CD18, anti-CD19 and anti-carcinoembryonic antigen (CEA) (Abcam, Cambridge, MA, USA). The sections were then incubated with Alexa Fluor 488-conjujated anti-mouse/anti-rabbit secondary antibodies (Cell Signaling Technology, Inc., Danvers, MA, USA), for 1 h at room temperature. Nuclei were stained with Hoechst 33342 (Invitrogen Life Technologies).

The SGECs on coverslips were fixed in 4% paraformaldehyde for 10 min, permeabilized with 0.1% Triton X-100 (Sigma) for 20 min at room temperature. Subsequently, 10% FBS was used to block non-specific binding. The SGECs were then incubated with primary antibodies at 4°C overnight. The antibodies that were used were the following: anti-CD7, anti-CD8, anti-CD14, anti-CD15, anti-CD18, anti-CD19, anti-CEA and anti-LGR5 (Abcam). The cells were then incubated with Alexa Fluor 488-conjujated anti-mouse/anti-rabbit secondary antibodies (Cell Signaling Technology, Inc.) for 1 h at room temperature. Nuclei were stained with Hoechst 33342.

Proteins (20 μg/lane) were fractionated by SDS-PAGE and electrotransferred onto polyvinylidene difluoride membranes. The blots were first incubated for 1 h in a blocking buffer consisting of 0.1% Tween-20 (Invitrogen Life Technologies) and 5% non-fat powdered milk, then incubated with a primary antibody at 4°C overnight. The antibodies that were used were as follows: anti-CD7, anti-CD8, anti-CD14, anti-CD15, anti-CD18, anti-CD19 (Abcam) and anti-β-actin (Cell Signaling Technology, Inc.). A horseradish peroxidase-conjugated, goat anti-mouse/anti-rabbit secondary antibody (Zhongshan Jinqiao Biotechnology Co., Ltd., Beijing, China) then was used and antigen-antibody complexes were detected by chemiluminescence using the BeyoECL Plus kit (BiYunTian Biotechnology Research Laboratory, Haimen, China).