Extended Data Fig. 2: VIL1 does not regulate the plant growth through PIF4.
Hypocotyl length of seedlings of each genotypes grown for 7 days at four different temperatures (12 °C, 17 °C, 22 °C, and 27 °C) in SD. b, Quantification of hypocotyl length, n = 20 seedlings. The box plot boundaries reflect the interquartile range, the horizontal line is the median and the whiskers represent 1.5X the interquartile range from the lower and upper quartiles. The letters marked as same color above each box indicate statistical difference between the genotypes at each temperature determined by One-way ANOVA followed by Tukey HSD test for multiple comparisons (P < 0.05). c and d, PIF4 mRNA expression in Col-0, vil1-1, phyB-9 and vil1-1phyB-9 seedlings grown for 7 days at four different temperatures in SD. Samples were collected at both ZT0 (c) and ZT6 (d). The expression levels were normalized to that of PP2A and further normalized by the transcript level of Col-0 at ZT0, at 12 °C. Error bars: ± s.d., n = 2, biological replicates (each biological replicate is an average value of four technical replicates). The letters marked as same color above each bar indicate statistical difference between the genotypes at each temperature determined by One-way ANOVA followed by Tukey HSD test for multiple comparisons (P < 0.05). e, Immunoblot analysis showing that endogenous PIF4 protein levels are not changed by vil1. Proteins were extracted from seven-day-old seedlings grown in SD at ZT6. Endogenous tubulin levels were detected as controls by anti-tubulin antibody. The experiment was repeated at least two times independently with similar results.
