Methyl viologen dichloride hydrate paraquat pq

All animals were grown at 20°C and cultured on NGM plates. The Bristol strain N2 was used as the wild type. The mutations used in this study are as follows: nuo-6(qm200), sod-2(ok1030), bli-3(e767), duox-2(ok1775) I, sod-1(tm783) II, isp-1(qm150), let-60(n1046), let-60(n1700), let-60(ga89) and lin-1(e1026) IV. For temperature shift experiments, let-60(ga89) adults were switch to the restrictive temperature (26°C) as adults and the progeny was used for the assay.
For testing the effects of Methyl viologen dichloride hydrate/Paraquat (PQ; Sigma-Aldrich 856177) and N-Acetyl-Cysteine (NAC; Sigma-Aldrich A7250), compounds were dissolved in water and stored at 4°C. The compounds were added to the NGM just before pouring the plates. The final concentrations of PQ and NAC used for vulva studies were 0.1 mM and 9 mM, respectively.

Benzoate, 3-hydroxyBenzoate, 4-hydroxyBenzoate, cinnamate, 3-hydroxycinnamate, salicylate, nicotinate, anthranilate, 4-aminoBenzoate, 2-aminophenol, benzamide, simazine, tryptophan, and methyl viologen dichloride hydrate (paraquat, PQ) were obtained from Sigma-Aldrich (St. Louis, MO, United States). Benzene (>99.7% purity), toluene, biphenyl, naphthalene, phenanthrene, anthracene, fluorene, and 30% (w v^-1) hydrogen peroxide (H₂O₂) were purchased from Merck (Darmstadt, Germany).

The required chemicals viz., Methyl viologen dichloride hydrate/Paraquat (PQ) (Sigma-Aldrich; St. Louis, MO, USA, Cat. No. 856177), Sucrose (SRL, Maharashtra, India, Cat. No. 1947139), DMSO (Sigma-Aldrich; St. Louis, MO, USA, Cat. No. D8418). Phosphate buffered Saline (PBS; HiMedia, Maharashtra, India, Cat. No. ML023), Trichloro Acetic Acid (TCA; SRL, Maharashtra, India, Cat. No. 204842), Dopamine (DA; Sigma-Aldrich, St. Louis, MO, USA, Cat. No. H8502); 3,4-Dihydroxyphenylacetic acid (DOPAC; Sigma-Aldrich, St. Louis, MO, USA, Cat. No. 11569); Homovanillic acid (HVA; Sigma-Aldrich, St. Louis, MO, USA, Cat. No. 69673) were used for quantifying DA and metabolites. Paraformaldehyde (Sigma-Aldrich, St. Louis, MO, USA, Cat. No. I58127), Triton X-100 (Sigma-Aldrich, St. Louis, MO, USA, Cat. No. T8787), Triton X-100 (Sigma, St. Louis, MO, USA), Normal goat serum (NGS, Vector labs, CA, USA), VECTASHIELD mounting medium (Vector Labs, CA, USA, Cat: H1000), Rabbit anti-Tyrosine hydroxylase (anti-TH) polyclonal primary antibody (Millipore, MA, USA, Cat: Ab152) and Goat anti-rabbit IgG H&L (TRITC labeled) polyclonal secondary antibody (Abcam, MA, USA, Cat: Ab6718) were used for immunostaining. Whatman filter paper no. 1 disk was used as a feeding medium in the experiment in a 30 × 100 mm glass vial.