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. 2003 Feb 1;31(3):935–943. doi: 10.1093/nar/gkg174

Figure 7.

Figure 7

zPER2 induces the cytoplasmic distribution of the zCLOCK1:zBMAL3 heterodimer. (A) Quantitative analysis of the sub- cellular localization of zCLOCK1 or zBMAL3 with or without zPER2 was done as described in Figure 4B. (B) Quantitative analysis of the sub- cellular localizations of zCLOCK1 and zBMAL3 expressed by pCLOCK1BMAL3 in the absence or presence of both zCRY1a and zPER2 was done as described in Figure 4B. VP16-tagged zCLOCK1 and FLAG-tagged zBMAL3 were stained as described in Figure 4A. HA- and V5-tagged zPER2 proteins, respectively, were stained with combinations of anti-HA antibody and FITC-conjugated secondary antibody and anti-V5 antibody and Cy3-conjugated secondary antibody. Nuclei were stained with DAPI. VP16-zCLOCK1, FLAG-zBMAL3, zCRY1a-HA and zPER2-V5 were expressed together in NIH3T3 cells by the co-transfection of pCLOCK1BMAL3 with pCRY1aPER2 vectors. Sub-cellular distributions were determined by double staining with a combination of zCLOCK1 (anti-VP16 antibody) and zPER2 (anti-V5 antibody) or with zBMAL3 (anti-FLAG antibody) and zCRY1a (anti-HA antibody).