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. 2003 Aug 1;17(15):1921–1932. doi: 10.1101/gad.1099503

Figure 2.

Figure 2.

Effects of CLOCK and BMAL1 coexpression on their posttranslational modifications. (A) Western blot analysis of HEK-293 cells transfected with different combinations of Clock and Bmal1 expression plasmids. Dose dependence of HA-BMAL1 posttranslational modification (B), and transactivation of mPer1-Luc reporter (C) on HA-CLOCK amount. HEK-293 cells were transfected with constant amounts of pcHA-Bmal1 (100 ng), mPer1-Luc (50 ng), and pcDNA3-lacZ and indicated amount of pcHA-Clock. Dose dependence of HA-CLOCK abundance (D) and transactivation of mPer1-Luc reporter (E) on HA-BMAL1 amount. HEK-293 cells were transfected with constant amounts of pcHA-Clock (300 ng), mPer1-Luc (50 ng), and pcDNA3-lacZ and indicated amounts of pcHA-Bmal1. Expression of HA-CLOCK and HA-BMAL1 proteins in transfected cells were detected by Western blotting with anti-HA antibodies. Typical result from three independent experiments is presented. Note that in cells transfected by pcHA-Clock only, anti-HA antibodies at longer exposures recognize nonspecific band with electrophoretic mobility close to HA-BMAL1.