Figure 7. Expression of
ipla-1 and
acl-10. (A-C) Confocal images of transgenic worms expressing
acl-10p::GFP. Seam cells (A-C: arrowheads), epithelial syncytium
hyp7 (A: bracket), vulval epithelium (B: arrows), and several neurons including AIYL/R (C: arrowheads) in the head region at the adult stage. Scale bars, 40 um. (D) Confocal images of epithelial cells of a transgenic L4 worm expressing IPLA-1::mCherry and ACL-10::GFP. Bar, 20 um. (E) Subcellular fractionation of IPLA-1 and ACL-10. Lysate of transgenic worms expressing ACL-10::mCherry and ACS-20::EGFP (
acs-20;
acs-22;xhEx3529[
acl-10p::
acl-10::mCherry]; tmEx1920[
acs-20p::
acs-20::egfp]) were subjected to S100 (cytosol)/P100 (membrane) fractionation and processed for immunoblotting with anti-IPLA, anti-mCherry (ACL-10), anti-{alpha}-tubulin (cytoplasmic marker), and anti-GFP (ER marker ACS-20) antibodies.
acs-20 encodes a very long chain fatty acid acyl-CoA synthetase that is known to be localized in ER membranes.
acs-22 is a homologous gene of
acs-20.
acs-20p::
acs-20::egfp transgene fully rescues the phenotypes of
acs-20;
acs-22 double mutants (Kage-Nakadai et al., 2010).