Fig 1: RUNX3 promoted osteogenic differentiation of ADSCs via upregulating hsa_circ_0005752. A. The mRNA levels of RUNX3 and hsa_circ_0005752 were determined by qRT-PCR in OM-induced ADSCs after transfection with pcDNA3.1 empty vector, pcDNA3.1-RUNX3 single or together with sh-NC or sh- hsa_circ_0005752. B-C. The osteogenic differentiation was assessed by the ALP staining (B), and ARS staining (C) in OM-induced ADSCs after transfection with pcDNA3.1 empty vector or pcDNA3.1-RUNX3 single or together with sh-NC or sh- hsa_circ_0005752. D. The protein levels (MDM2, p53, Runx2, ALP, Osx, and OCN) were measured by western blot in OM-induced ADSCs after transfection with pcDNA3.1 empty vector, pcDNA3.1-RUNX3 single or together with sh-NC or sh- hsa_circ_0005752. Data indicated the mean ± SD, n = 3. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.01.
Fig 2: RUNX3 regulated hsa_circ_0005752 expression during ADSCs osteogenic differentiation. A-B. The RUNX3 mRNA or protein levels was detected in ADSCs after osteogenic differentiation for 0, 3, 7, and 14 days by qRT-PCR (A) or western blot (B). C. The binding capacity of RUNX3 to LAPR1 promoter was validated by the CHIP-qPCR experiment in ADSCs. D. The effects of RUNX3 on LAPR1 transcription were detected by luciferase reporter assay. E. The LPAR1 protein levels were detected in ADSCs after overexpressing or knockdown RUNX3 by western blot. F. The hsa_circ_0005752 levels were detected in ADSCs after overexpressing or knockdown RUNX3 by qRT-PCR. Data indicated the mean ± SD, n = 3. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.01.
Fig 3: RUNX3 promotes the expression of circDYRK1A and inhibits the progression of GC. A The expression of RUNX3 in microarray dataset GSE49051. B The expression of RUNX3 in GC tissues and adjacent normal tissues detected by RT-qPCR (n = 50). C RUNX3 expression in GC tissues and adjacent normal tissues measured by immunohistochemistry. D Pearson correlation analysis of the correlation between RUNX3 expression and circDYRK1A expression in GC tissues. E The enrichment of RUNX3 in circDYRK1A promoter region detected by ChIP assay. F The effect of RUNX3 on circDYRK1A promoter activity. G Expression of RUNX3 and circDYRK1A in AGS and HGC-27 cells measured by dual luciferase reporter assay. H Tumor size (left panel), growth curve (middle panel) and weight (right panel) in each group of mice detected (n = 6). I The apoptosis of tumor cells detected by TUNEL staining. Cell nuclei were stained in blue by hematoxylin, and yellow–brown cells are positive cells. J The expression of GLS and GDH (n = 6) measured by immunohistochemistry. The cell experiment was repeated three times. * p < 0.05
Fig 4: Graphical summary of mechanism of circDYRK1A in GC. RUNX3 enhances the expression of circDYRK1A, which in turn acts as a miR-889-3p sponge and upregulates FBXO4 expression, thus inhibiting glutamine metabolism and malignant phenotypes of GC cells to repress the progression of CG
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