Heat exposure began each day of the stress treatment with an increasing temperature gradient from 15 °C to 45 °C over a period of 5 h, and this was then maintained at 45 °C for 6 h. During the following 5 h, the temperature was returned to 15 ºC and maintained for 8 h, thus mimicking a day-night scenario. Control plants (C) were sampled, and the following day the stress exposure began. Plant material was sampled at the end of the 6 h heat exposure on day 1 (T1), day 3 (T3), and day 5 (T5). To characterize the molecular mechanisms driving heat response, we accomplished a systems biology analysis combining two subcellular bottom-up proteomics assays.