FIG. 4.

Transcriptional changes in Wnt signaling and Clock genes after Rev-erbα overexpression and addition of Wnt3a protein. (A) Differences in Rev-erbα mRNA levels among the groups. The transcriptional levels of Rev-erbα decreased when Wnt3a was added (Rev-erbα +Wt3a group). (B) Differences in Bmal1 mRNA levels among the groups. The trend was nearly opposite that observed for Rev-erbα. (C) The transcriptional level of Wnt3a among the groups. Less Wnt3a mRNA was expressed in the Rev-erbα group. (D) The transcriptional level of β-catenin among the groups. The least expression was observed in the Rev-erbα group, and the most expression was seen in the Wnt3a group. (E) The expression of T cell factor 1 (TCF1) mRNA among groups. The expression was decreased following Rev-erbα overexpression and was increased when Wnt3a protein was added. (F) The expression of glycogen synthase kinase (GSK)-3β mRNA among groups. The Rev-erbα mRNA expression trend was nearly unchanged. (G) Western blot results of Rev-erbα protein changes among the groups. The trend was almost the same as the Rev-erbα mRNA expression trend. Rev-erbα protein levels for the (I) Rev-erbα, (II) Rev-erbα +Wnt3a, (III) EGFP, (IV) EGFP+Wnt3a, (V) control, and (VI) Wnt3a groups. (H) Quantified using densitometry by Quantity One software, the ratio of Rev-erbα relative to GAPDH is expressed as the relative content of Rev-erbα protein. Data represent the mean±SD (n=3), and the * symbol denotes a significant difference (P<0.05). The # symbol indicates P<0.05 compared with the other groups.