Molecular Mechanism of Long Noncoding RNA SNHG14 in Osteogenic Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells through the NEDD4L/FOXA2/PCP4 Axis

Figure 5

NEDD4L inhibits FOXA2 expression through ubiquitination modification and promotes PCP4 transcription. (a) Binding of NEDD4L to FOXA2 was examined by coimmunoprecipitation. (b) FOXA2 expression level after osteogenic induction (OI) was determined by Western blot analysis. (c) FOXA2 expression level in BMSCs was determined by Western blot analysis. (d) Ubiquitination level of FOXA2 was determined by the ubiquitination assay. (e) Binding of FOXA2 to the PCP4 promoter was predicted on the JASPAR database. (f, g) FOXA2 to the PCP4 promoter was analyzed by dual-luciferase and ChIP assays. (h, i) mRNA level of PCP4 after OI and in BMSCs was determined by RT-qPCR. Cell experiments were repeated 3 times. Data were represented as deviation. Multigroup comparisons in panels (b) and (c) and (h) and (i) were analyzed using one-way ANOVA and in panels (f) and (g) were analyzed using two-way ANOVA, followed by Tukey’s multiple comparison test. , .

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