Cfr5000

For analysis of GFP, mCherry or SNB-1::GFP, live animals were anesthetized using 1% (v/v) 1-phenoxy-2-propanol in M9 buffer and visualized using a 40× magnification oil-immersion lens and an epifluorescent microscope (Leica CFR5000). A CCD camera (Leica DFC345 FX) was used for documentation. Images were analyzed and scale bars applied using Leica Application Suite for Advanced Fluorescence (LAS-AF) software. Termination defects were quantified by manually scoring juIs76 (P_unc-25::GFP) for dorsal cord tips, and bggIs6 (P_unc-25::mCherry) for DD5. Each genotype was quantitated by manually scoring three counts of 10 to 20 animals from two or more independent experiments.

For analysis of RPM-1::GFP, live adult animals were anesthetized using Levamisole in M9 buffer and visualized using an epifluorescent microscope (Leica CFR5000) with a 40× magnification oil-immersion lens. Images were acquired using a CCD camera (Leica DFC345 FX). Ten animals per strain were observed across two independent days with representative images shown in Figure 5.