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. Author manuscript; available in PMC: 2013 Apr 29.
Published in final edited form as: J Neurosci. 2012 Apr 4;32(14):4867–4877. doi: 10.1523/JNEUROSCI.5650-11.2012

Figure 1. Light induces the phosphorylation of RKIP and its dissociation from c-Raf in the SCN.

Figure 1

A, Representative micrographs of total RKIP expression in the murine SCN at ZT 8. Boxed region (left) is shown in high magnification on the right. B, Light-responsive interaction between RKIP and c-Raf in the SCN as determined by co-IP. Mice received a 15 min light pulse (LP: 400 lux intensity) at CT 15 and were killed 5 min later. Control mice were killed at the same circadian time without receiving a light pulse (DD). SCN protein extracts were immunoprecipitated (IP) with antibodies against total RKIP (top) or total c-Raf (bottom) and immunoblotted (IB) with both antibodies. IP with rabbit IgG was used as a negative control. A brief light pulse in the early subjective night reduces the binding interaction of RKIP and c-Raf. C, Light-induced phosphorylation of RKIP as determined by Western blot analysis. Mice received a 15 min light pulse (LP: 400 lux intensity) at CT 15 and were killed 5 min later. Control mice were killed at the same circadian time without receiving a light pulse (DD). Blots were probed with antibodies against the Thr-153-phosphorylated form of RKIP (p-RKIP) (top), total RKIP (middle), and total c-Raf (bottom). Light triggers rapid and robust phosphorylation of RKIP at Thr-153 in the SCN.