Figure 8.

Polysome Loading Analysis.

Polysome gradients were performed for Col-0 plants grown in five different thermocycles, harvested at ED (white bars) and EN (dark-gray bars).

(A) Examples of the distribution of ribosomes in fractions collected from a density gradient obtained from plants harvested at EN and grown in a 24°C/24°C (left panel) and 24°C/12°C (right panel) thermocycle. RNA was measured as absorbance at 254 nm (A₂₅₄). Free ribosomes and monosomes are on the left-hand side and increasingly large polysomes toward the right-hand side of the display.

(B) Estimated proportion of ribosomes in polysomes. This is calculated as (PS)/(NP + PS). The percentage is given as numbers in the figure panel. The results are the mean ± sd (n = 3), except in the case of 12°C/12°C at EN, where n = 1.

(C) Ribosome content, estimated from the sum of the ribosome profile at A₂₅₄. The average of the ribosome number at ED and EN was calculated for each treatment. The results are given as mean ± sd (n = 4). One-way ANOVA was used to identify potential candidates for a statistically significant difference between treatments. After ANOVA P value correction using Holm’s method (P < 0.05), individual contrasts were then identified in a post-hoc Tukey HSD test (P < 0.05). They are indicated by different letters within the same time point (ED, lowercase; EN, uppercase). In the case of 12°C/12°C at EN, this analysis was not possible due to lack of replication (n = 1). Comparisons between ED and EN using a paired t test at a given thermocycle are indicated by an asterisk if significant (P < 0.05).