Ab109443

The paraffin-embedded sections (4 μm) of the spinal cords were mounted on glass slides. IHC analysis of the sections was performed using an automated slide preparation system (Benchmark XT; Ventana Medical Systems Inc., Tucson, AZ, USA). Deparaffinization, epitope retrieval, and immunostaining were performed using cell conditioning solutions and the BMK ultraVIEW diaminobenzidine detection system (Ventana Medical Systems Inc., Tucson, AZ), following the manufacturer’s instructions. The tissue sections were stained with anti-CD3 (#ab16669, Abcam), anti-CD68 (#ab125212, Abcam), and anti-PAX5 (#ab109443, Abcam) primary antibodies. The positive signals were amplified using ultraVIEW copper. The sections were counterstained with hematoxylin and bluing reagent. The number of CD3-, CD68-, and PAX5-immunoreactive cells in the white matter and pia mater of the spinal cord was examined using a light microscope and normalized to the total area of white matter and pia mater.

Total proteins were extracted from indicated CC cells using RIPA buffer, and then isolated by 10% SDS‐PAGE. Subsequently, the target proteins were transferred into PVDF membranes followed by the incubation of non‐fat dry milk (5%) for 2 hours. Next, the membranes were probed by primary antibodies that against PAX5 (1:5000, ab109443, Abcam) and β‐catenin (1:5000, #9581, Cell Signaling Technology) for 24 hours, followed by secondary antibody incubation for 2 hours. The bands were visualized by chemiluminescence.