Expr16527
UNC-87 and CLIK-1 are segregated into different subdomains in sarcomeric actin filaments in the body wall muscle. In immunofluorescence microscopy using anti-UNC-87 antibody, UNC-87 co-localized with actin in sarcomeres. In our immunofluorescent staining for actin using anti-actin monoclonal or polyclonal antibody, a portion of sarcomeric actin near the pointed ends are not stained well as reported previously (S. Ono et al., 2022). Therefore, closely matched patterns of UNC-87 and actin in immunostaining indicated that UNC-87 was also absent from the edges of sarcomeric actin where the pointed ends of actin filaments were concentrated. In contrast to UNC-87, CLIK-1::GFP was concentrated in sharp lines at the edges of phalloidin-stained sarcomeric actin filaments. A similar localization pattern of CLIK-1::GFP was also observed in live worms without fixation, and its concentration to the edges of sarcomeric actin bands was confirmed by comparing with the pattern of mCherry::LifeAct, indicating that the localization of CLIK-1::GFP was not an artifact of fixation. Furthermore, CLIK-1::GFP was not colocalized with ATN-1 α-actinin, which is concentrated at the dense bodies near the barbed ends of actin filaments, suggesting that CLIK-1::GFP localized near the pointed ends of actin filaments. These results demonstrate that the two calponin-related proteins, CLIK-1 and UNC-87, are segregated within sarcomeric actin filaments in C. elegans striated muscle. Since CLIK-1::GFP was localized near the pointed ends of sarcomeric actin filaments, the location of CLIK-1::GFP was compared with that of UNC-94 tropomodulin that localizes to the pointed ends of sarcomeric actin filaments in the C. elegans body wall muscle (Stevenson et al., 2007; Yamashiro et al., 2008). Comparison of the locations of CLIK-1::GFP and UNC-94 indicated that an overlap between these two proteins was minimum, suggesting that CLIK-1 is enriched near the pointed ends of sarcomeric actin filaments but not extended to the pointed ends.
Expr16528
UNC-87 and CLIK-1 are segregated into different subdomains in sarcomeric actin filaments in the body wall muscle. In immunofluorescence microscopy using anti-UNC-87 antibody, UNC-87 co-localized with actin in sarcomeres. In our immunofluorescent staining for actin using anti-actin monoclonal or polyclonal antibody, a portion of sarcomeric actin near the pointed ends are not stained well as reported previously (S. Ono et al., 2022). Therefore, closely matched patterns of UNC-87 and actin in immunostaining indicated that UNC-87 was also absent from the edges of sarcomeric actin where the pointed ends of actin filaments were concentrated. In contrast to UNC-87, CLIK-1::GFP was concentrated in sharp lines at the edges of phalloidin-stained sarcomeric actin filaments. A similar localization pattern of CLIK-1::GFP was also observed in live worms without fixation, and its concentration to the edges of sarcomeric actin bands was confirmed by comparing with the pattern of mCherry::LifeAct, indicating that the localization of CLIK-1::GFP was not an artifact of fixation. Furthermore, CLIK-1::GFP was not colocalized with ATN-1 α-actinin, which is concentrated at the dense bodies near the barbed ends of actin filaments, suggesting that CLIK-1::GFP localized near the pointed ends of actin filaments. These results demonstrate that the two calponin-related proteins, CLIK-1 and UNC-87, are segregated within sarcomeric actin filaments in C. elegans striated muscle. Since CLIK-1::GFP was localized near the pointed ends of sarcomeric actin filaments, the location of CLIK-1::GFP was compared with that of UNC-94 tropomodulin that localizes to the pointed ends of sarcomeric actin filaments in the C. elegans body wall muscle (Stevenson et al., 2007; Yamashiro et al., 2008). Comparison of the locations of CLIK-1::GFP and UNC-94 indicated that an overlap between these two proteins was minimum, suggesting that CLIK-1 is enriched near the pointed ends of sarcomeric actin filaments but not extended to the pointed ends.