Fig. 5: BCY and YHB-YFP display distinct temperature-insensitive PB patterns. | Nature Communications

Fig. 5: BCY and YHB-YFP display distinct temperature-insensitive PB patterns.

From: Increasing ambient temperature progressively disassembles Arabidopsis phytochrome B from individual photobodies with distinct thermostabilities

Fig. 5

a Maximum-projection, deconvolved images showing representative steady-state patterns of phyB C-terminal module fused with YFP (BCY) or YHB-YFP PBs in cotyledon epidermal-cell nuclei of 4-day-old BCY and YHB seedlings, respectively, grown at 12, 16, 21, and 27 °C in the dark or in 10 µmol m−2 s−1 in R light. Nuclei were labeled by DAPI (blue), white arrowheads indicate Nuo-PB, and the boundaries of the nucleoli are traced by red dashed lines. The percentage of nuclei with s.e. showing the representative PB pattern is shown in each image; n indicates the total number of nuclei analyzed. Scale bars represent 5 μm. b, c Quantification of the numbers of total, Nuo, and nonNuo PBs per nucleus in cotyledon epidermal-cell nuclei of the BCY (b) and YHB (c) seedlings described in a. d Quantification of the percentage of nuclei with the indicated number of PBs in cotyledon epidermal-cell nuclei of the BCY and YHB seedlings described in a. Error bars represent s.e. calculated from groups of seedlings. Different letters denote statistically significant differences (ANOVA, Tukey’s HSD, P ≤ 0.05, n = 3). e Cotyledon size measurements of the BCY or YHB seedlings described in a. f Immunoblot results showing the levels of BCY or YHB-YFP in 4-day-old BCY or YHB seedlings grown in the dark or 10 µmol m−2 s−1 R light in the indicated temperatures. BCY and YHB-YFP were detected by anti-phyB antibodies. RPN6 was used as a loading control. The relative levels of BCY and YHB-YFP normalized against the corresponding levels of RPN6 are shown. g Quantification of the volume of BCY and YHB-YFP PBs in cotyledon epidermal-cell nuclei of the BCY and YHB seedlings described in a. For b, c, e, g, the boxes represent from the 25th to the 75th percentile and the bars equal the median values. Samples with different letters exhibit statistically significant differences (ANOVA, Tukey’s HSD, P ≤ 0.05, for b, c, and g, n ≥ 9, for e, n ≥ 78). The source data underlying the PB measurements in b–d, g, cotyledon measurements in e, and immunoblots in f are provided in the Source Data file.

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