Levamisol hydrochloride

Synchronized animals were treated with 60 µg/ml cisplatin for 24 h. Then, levamisol hydrochloride (Sigma-Aldrich, 31742)-anesthetized animals were mounted on a microscope slide and covered with a cover slip. Worms were pictured using an Imager2 Zeiss fluorescence microscope, and the same exposure time was applied to all experimental conditions. GFP expression was quantified using ImageJ software, measuring the fluorescence intensity of the whole animal. Three experimental replicates were performed. For each condition, at least 20 worms were acquired and quantified.

For quantification of hsp-6::GFP expression, animals, exposed or not to 100 µg/ml cisplatin from L4/YA stage for 24 h, were transferred to a microscope slide and anesthetized with 15 µl of 10 mM levamisol hydrochloride (Sigma-Aldrich, 31742) dissolved in S-Basal [5.85 g NaCl, 1 g K₂HPO₄, 6 g KH₂PO₄, 1 ml cholesterol (5 mg/ml in ethanol), H₂O to 1 l], and sealed with a cover slip. Worms were pictured at 25× magnification using an Imager2 Zeiss fluorescence microscope, and the same exposure time was applied to all experimental conditions. GFP expression was quantified using ImageJ software (https://imagej.nih.gov/ij/), measuring the fluorescence intensity of the pharyngeal bulb region of each worm acquired. Three experimental replicates were performed, and, for each condition, 10-15 worms were pictured and quantified.