Figure 2. HNF4α2 is a major regulator of osteogenesis and metabolism in osteoblasts.

(A and B) Hnf4α1/2 mRNA expression in differentiated primary bone marrow stromal cells (BMSCs), mature osteoblasts (OBs), and MC3T3-E1 osteoblasts (A), and in MC3T3-E1 osteoblasts transfected with an empty vector (Ctr), Hnf4α1 (Hnf4α1^Tg), and Hnf4α2 (Hnf4α2^Tg) expression transgene (B). (C and D) mRNA expression of markers of osteoblast differentiation Runx2 and Sp7. Values are expressed as the mean ± SEM. n ≥ 3 per group of a representative experiment performed at least 3 times; corrected P < 0.05 versus *BMSC or Ctr. Statistical analysis was performed with an ANOVA test followed by post hoc t tests to determine statistical differences and multiple-testing correction using the Holm-Bonferroni method. (E) Number of differentially regulated genes identified by RNA-Seq in Hnf4α1^Tg and Hnf4α2^Tg osteoblasts versus Ctr. (F) Canonical pathway analysis and prediction of pathway activation of differentially regulated genes identified by RNA-Seq of Ctr, Hnf4α1^Tg, and Hnf4α2^Tg osteoblasts. (G and H) Heatmap-represented expression of genes modified and involved in osteogenesis and metabolism pathways in Ctr, Hnf4α1^Tg, and Hnf4α2^Tg osteoblasts. Corrected P < 0.05; n = 3 per group. Statistical analysis was performed with an unpaired Student’s t test and corrected by the FDR.