Expr2560
CLC-2-GFP was detected in hypodermal seam cells, which fuse to form a single elongated syncytium. However, in contrast to CLC-1-GFP, under the expression condition used in this study, GFP signals did not appear to be concentrated at the borders between the seam cell syncytium and surrounding hypodermal cells. When adult wild-type worms were whole-mount stained with anti-CLC-2 pAb, the seam cell syncytium appeared to be outlined to give two parallel lines, although these lines were thick and not very sharp.
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.
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.