Visualizing unc-49 Expression in AVA Neurons

To test whether unc-49 is expressed in AVA interneurons, we coinjected four plasmids, wp1427 (Punc-49::GFP), wp1339 (Pgpa-14::Cre), wp1392 (Pflp-18::loxP::LacZ::STOP::loxP::mStrawberry), and lin-15(+) into the lin-15(n765) strain. The expression of wp1339 and wp1392 results in mStrawberry labeling of the two AVA interneurons^{34 (link),81 (link)}. The expression patterns of GFP and mStrawberry were imaged with an inverted microscope (TE-2000U, Nikon) enhanced GFP/fluorescein isothiocyanate and mCherry/Texas Red filter sets (49002 and 49008, Chroma Technology Corporation, Rockingham, VT, USA) and a Hamamatsu ORCA-Flash4.0 digital camera (Model C11440-22CU). Pflp-18 and Pgpa-14 were gifts from Dr Alexander Gottschalk^{81 (link)} whereas Punc-49 was cloned from genomic DNA of the Bristol N2 strain by PCR using primers listed in Supplementary Table 1.