Fig. 4 |. Experimental designs for analysing responses to ER stress in Arabidopsis.

a, Adaptive endoplasmic reticulum (ER) stress. In this approach, 5-day-old seedlings are subjected to a high dose of tunicamycin (Tm) (for example, 500 ng ml⁻¹) in liquid growth media using multi-well plates for a duration determined by the experimental objectives. The Tm solvent dimethyl sulfoxide (DMSO) is used as mock control. b, Chronic ER stress. Seeds are directly sown onto solid growth media containing a low dose of Tm (for example, 25 ng ml⁻¹) or DMSO as control. Different doses of Tm can induce various cellular environments (for example, low dose for pro-survival environment; high dose for pro-death environment) that researchers can utilize to investigate their biological questions. Bi-plates featuring Tm-containing media on one side and DMSO-containing media on the other are advantageous for this method. Phenotypic assessments are typically conducted between 9 and 14 days after planting. c, ER stress recovery. After exposure to adaptive ER stress, seedlings are transferred to plates containing solid growth media for recovery. Growth restoration is evaluated between 5 and 7 days after the transfer. Collectively, these experimental systems offer powerful means to analyse molecular and physiological responses to ER stress in a highly reproducible manner.