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. 2007 Apr 23;177(2):205–210. doi: 10.1083/jcb.200607084

Figure 2.

Figure 2.

Overexpression of UNC-45FLAG results in diminished accumulation of body wall myosins. (A) C. elegans worms overexpressing UNC-45FLAG or harboring an UNC-45 Lof mutation have decreased levels of body wall muscle MHC. Individual worms from N2, suIs2, and unc-45(e286) strains at either the permissive (15°C) or restrictive (25°C) temperatures were isolated at the Ya stage. Western blots against MHC A, B, and D show that only body wall muscle MHC A and B are affected in transgenic UNC-45FLAG worms, whereas MHC D is also decreased in e286 mutant worms. (B) Quantification of MHC levels in worms overexpressing UNC-45FLAG and in UNC-45 Lof mutants. Analysis of Western blots performed in triplicate show that suIs2 worms have 30% less MHC A and B than N2 wild-type worms. Note that this decrease is not as great as that of e286 mutants, which have 70% less body wall MHC than wild type. *, P < 0.5; **, P < 0.01, versus N2 wild type. (C) MHC mRNA levels between N2 and suIs2 remain unchanged. The threshold cycle (Ct) was set as the fractional cycle number at which the real-time fluorescence signal of the reaction could be measured above background. The cycle at which N2 wild type and suIs2 crossed the threshold was within one cycle each for unc-54, myo-3, and GAPDH. Error bars indicate SD.