Fig. 2

The expression of ITGBL1 in hepatocytes is regulated by RUNX2. (A) The mRNA expression data from liver biopsies of CHB patients in the GSE84044 cohort were analyzed using Pearson correlation coefficients to assess the relationship between ITGBL1 and RUNX2. A significant positive correlation was observed between ITGBL1 and RUNX2 (Pearson’s r = 0.66603, p < 0.01). (B) Huh7 cells seeded in 6-well plates were transfected with RUNX2 overexpression plasmid, shRUNX2 plasmid, and shscramble plasmid as a negative control, with untreated cells as control. Cells were harvested 48 h post-transfection, and Western Blot was used to analyze the expression of ITGBL1 and RUNX2. In Huh7 cells, the expression of ITGBL1 was enhanced with RUNX2 overexpression and downregulated with RUNX2 knockdown. (C) ChIP experiments in Huh7 cells showed that RUNX2 binds to the ITGBL1 promoter. (D) The ITGBL1 promoter plasmid and shRUNX2 plasmid were co-transfected into HEK293T cells (96-well plate), and the transcriptional activity of the ITGBL1 promoter was detected by dual-luciferase reporter assays 48 h after co-transfection. The activity of the ITGBL1 promoter increased with RUNX2 overexpression and decreased with RUNX2 knockdown. (E) Huh7 cells seeded in 6-well plates were treated with increasing concentrations of the RUNX2 inhibitor, Vitamin D3. After 48 h of treatment with Vitamin D3, Western Blot was used to detect the expression of ITGBL1 and RUNX2. As the concentration of Vitamin D3 increased, RUNX2 expression gradually decreased, and the expression of ITGBL1 was downregulated in accordance with the reduction in RUNX2 expression. The figure shows a representative image