SQST-1:: and ::LGG-1 CemOrange2 fusion proteins monitor autophagic flux.

Pnhx-2sqst-1::CemOrange2 (A, C, E; red) and Pnhx-2CemOrange2::lgg-1 (B, D, F; red) transgenic animals were treated with a diluent control (0.1% DMSO; A and B), 3-methyladenine (10 mM 3-MA, C and D), or 25 μM fluphenazine (E and F) for 16 hrs in liquid culture and analyzed using confocal microscopy over ≥ 20 z-planes Ex555 nm/Em565-590 nm. Representative confocal images of animals shown as a single XY plane. Scale bar = 25 μm. Multiple confocal images of transgenic animals (n ≥ 5 animals) expressing SQST-1::CemOrange2 (G) or CemOrange2::LGG-1 (H) after treatment with the above compounds, were quantified for number of puncta per μm2 imaged using the Quantification module in the Volocity image analysis software (v6.3). Statistical analysis of the drug-treated animals relative to diluent control was performed using an unpaired, 2-tailed t-test (*p<0.05).