Ab231345

The tissues were fixed in 4% paraformaldehyde and embedded in paraffin. Then, 5-μm sections were cut using a microtome (Leica Microsystems, Germany), dewaxed, dehydrated, and subjected to antigen retrieval. Subsequently, endogenous peroxidase was removed using 3% H₂O₂, and the tissue sections were incubated overnight at 4°C with primary antibodies against LRRC15 (1:200, M041540; Abmart Shanghai, China), CBF-β (1:200, ab231345; Abcam), and RUNX1 (1:1,000, PA5-85543; Thermo Fisher Scientific). The sections were incubated with HRP-coupled secondary IgG antibody (1:2,000, ab6721; Abcam) for 60 min at room temperature and stained with DAB reagent. After counterstaining with hematoxylin, the sections were photographed under a microscope and the tan-positive stained areas of each section were analyzed using ImageJ.
For H&E and Safranin O-fast green staining, paraffin-embedded sections were dewaxed, sequentially dehydrated, and stained using standard protocols (Beijing Solarbio Life Sciences, China). Finally, photographs were obtained under a light microscope to analyze pathological changes involving the mouse tissues.

Total protein was extracted from cells using RIPA Lysis Buffer (Phygene, China) and quantified using the BCA Protein Assay Kit (Phygene). Proteins were separated using 12% SDS-PAGE and transferred to PVDF membranes. The membranes were blocked with 5% bovine serum albumin for 2 h at room temperature and incubated overnight at 4°C with primary antibodies against LRRC15 (1:1,000, ab150376; Abcam, UK), RUNX1 (1:1,000, PA5-85543; Thermo Fisher Scientific, USA), CBF-β (1:1,000, ab231345; Abcam), and GAPDH (1:2,000, #5174; Cell Signaling Technologies, USA). After that, the membranes were further incubated with horseradish peroxidase (HRP)-coupled secondary IgG antibody (1:10,000, ab6721; Abcam) for 120 min at room temperature. Protein bands were examined using an ECL substrate kit (Abcam), and all proteins were quantified based on the internal reference band GAPDH using ImageJ software.