Fig. 6

Effect of shBRD4, rWNT4 and NF-κB inhibitors on glycolysis and osteogenic differentiation of hBMSCs. hBMSCs were transfected with shNC or shBRD4 and treated with rWNT4 or Bardoxolone Methyl followed by induced osteogenic differentiation. A Detection of WNT4, IκBα, p-IκBα, p65 and p-p65 expression using Western blot. B-D Glycolysis was detected by measuring glucose uptake, lactic acid production and ATP levels. E Western blot assay was used to examine the expression levels of key glycolytic enzymes (HK2, LDHA, PKM2 and PFK1). F ALP activity was tested by ALP Assay Kit. G ARS was utilized to assess mineralized deposit. H The protein expression levels of OCN, RUNX2, BSP and ALP were determined using Western blot analysis. n = 3; *P < 0.05, **P < 0.0, ***P < 0.001