To analyse the protein accumulation in different temperature and time, total proteins of 15 DAE NIL‐E1 PHYA2 PHYA3 plants from 25 and 35 °C were extracted with protein extraction buffer (50 mm Tris–HCl pH7.5, 150 mm NaCl, 5 mm EDTA, 0.1% Triton X‐100 and protease inhibitor cocktail). The homogenate was clarified by centrifugation at 12 000 g for 10 min at 4 °C, and aliquots of the supernatant were combined with 2× SDS sample loading buffer and heated at 95 °C for 5 min to denature the protein. The antibody anti‐phyA3 and anti‐phyA2 were previously reported (Lin et al., 2022 (link)), and anti‐actin was obtained from CWBIO (CW0264).