5 hydroxyindole 3 acetic acid

Chondroitin sulfate type A (CS A; sodium salt from bovine trachea; 70% with counterbalance CS type C [CS C]), CS C (sodium salt from shark cartilage; 90% with counterbalance CS A), alginic acid, sodium salt (Algin^®, sodium alginate), ι-carrageenan (commercial grade, type II), fucoidan (FUCO; from Fucus vesiculosus), serotonin.HCl, 5-hydroxyindole and 5-hydroxyindole-3-acetic acid were obtained from Sigma-Aldrich (WI, USA). CuCl₂.2H₂O was obtained from Fisher Scientific (GA, USA). All other reagents were of analytical grade.

All standards, including amino acids, salicylic acid, phenols, IAA, 5-Hydroxyindole-3-acetic acid, Indole-3-acetamide, N-(3-indoley lacetyl)-L-alanine, Indole-3-carboxaldehyde, Methyl indole-acetate, jasmonic acid (JA), abscisic acid, salicylic acid glucoside ester, OPDA, carboxylic acids, and sugars were purchased from SIGMA (Barcelona, Spain). Methanol (HPLC grade) was obtained in SIGMA (Barcelona, Spain), formic acid and NaOH were obtained from J.T Baker (Deventer, Holland). Indole-3-carboxylic acid and 1,4-diaminobutane were obtained from VWR (Barcelona, Spain).

RNAi experiments were conducted by feeding L4 staged animals on HT115(DE3) bacteria expressing dsRNA of the relevant target gene (chk-1) for 48 h until dissection.
NGM plates containing 1-mM 5-Hydroxyindole-3-acetic acid (Sigma) dissolved in absolute ethanol (+auxin) were used for auxin-induced degradation of cdk-1, cdk-2, and chk-2. The control plates (-auxin) were poured in the same way with the addition of an identical volume of ethanol without auxin. Given that presence of auxin inhibits bacterial growth, a saturated OP50 culture was concentrated 5× before being spotted onto NGM plates. Plates were left to dry overnight at room temperature before L4 animals were plated and then dissected 24 h later.