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. 2017 Dec 11;6:e24779. doi: 10.7554/eLife.24779

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© 2017, Hayasaka et al

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

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Figure 4. — (a) MEFs were established from Sik3^+/+ (WT) Per2^Luc/+ mice and Sik3^-/- (KO); Per2^Luc/+ mice. The cellular rhythms were synchronized by dexamethasone (DEX, 100 nM) treatment to examine rhythmic expression by western blotting analysis. PER2 (bottom band) and PER2::LUC (upper bands) expression levels at all time points in knockout (KO) mouse embryonic fibroblast (MEFs) are higher than those in wild-type (WT) MEFs. (b) Western blotting performed on liver nuclear extracts, Sik3^+/- (He); Per2^Luc/+ mice, and littermate Sik3^-/- (KO); Per2^Luc/+ mice. PER2 expression in liver nucleus was higher in Sik3^-/- mouse as compared to in Sik3^+/- mouse ZT22, left). To confirm the protein mobility of endogenous PER2, rhythmic expression of PER2 in Sik3^+/+ (±) mice was also detected. Note that Sik3^+/+ (WT) mice do not have Per2^Luc allele. Therefore, PER2 bands of Sik3 He and KO are from a single Per2 allele while those from WT are from both alleles.