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. 2011 Sep 15;124(18):3164–3173. doi: 10.1242/jcs.087320

Fig. 2.

Fig. 2.

Reduction in wild-type nematode motility, thick filament assembly and body-wall muscle myosin accumulation by transgenic UCS-containing proteins. (A) The UCS domain and TPR(−) constructs significantly decrease wild-type nematode motility. Error bars indicate mean ± s.d. ***P<0.001. (B) The UCS domain and TPR(−) construct significantly diminish A-band assembly in wild-type nematode. (C) Quantification of A-band assembly diminution of transgenic wild type. Error bars indicate mean ± s.d. ***P<0.001. N.S., not significant. (D) The UCS domain is expressed at low levels compared with other transgenic proteins and the myosin-binding truncates show reduction in body-wall muscle myosin accumulation similar to FL UNC-45 in wild-type nematodes. Transgenic expression of UNC-45 fragments was detected by immunoblots with anti-FLAG antibody. UCS was almost invisible by anti-FLAG antibody. Body-wall muscle-specific myosin heavy chains A and B were detected and quantified by immunoblots with mAb 5–6 and mAb 28.2. Pharyngeal myosin heavy chain D, detected by mAb 5–17, was used as loading control. (E) UNC-45 truncate expression in wild type was examined by immunoblots with rabbit polyclonal anti-C. elegans UNC-45 antibody. UCS and other fragments were detectable on the blots. Endogenous UNC-45 and FLAG-tagged full-length UNC-45 overlapped on the top of the immunoblot. Pharyngeal myosin heavy chain D was used as loading control. (F) A titration experiment of transgenic UCS worms in wild type was performed to semi-quantitatively compare the level of UCS expression with that of FL in FL transgenic wild type by rabbit polyclonal anti-C. elegans UNC-45 antibody. Endogenous UNC-45 and FLAG-tagged full-length UNC-45 overlapped on the top of the immunoblot. UCS became detectable by polyclonal anti-C. elegans UNC-45 antibody. Pharyngeal myosin heavy chain D was used as loading control. (G) Table 1 shows the ratio of protein amounts. Table 2 shows ratios of the decreased body bends and the diminished A-bands between transgenic FL and UCS worms compared with those of wild type.