Fig. 6: AHL22 cooperates with HDA15 to repress the expression of auxin responsive genes.
From: Chromatin attachment to the nuclear matrix represses hypocotyl elongation in Arabidopsis thaliana
a The hypocotyl phenotype of hda15-1 grown vertically under LD conditions (16 h at 23 °C under 22 μmol·m−2·s−1 continuous white light, 8 h at 23 °C dark) for 5 days. Scale bar= 0.5 cm. b Hypocotyl lengths of Col-0 and hda15-1. Average length of three independent measurements ± standard deviations are shown. Each measurement with n = 30 plants. Unpaired two-tailed Student’s t-test was used. N.S. p value > 0.05, *p value ≤ 0.05, **p value ≤ 0.01, ***p value ≤ = 0.001, ****p value ≤ = 0.0001. c Bimolecular fluorescence complementation (BiFC) analysis of AHL22 and HDA15 interaction in vivo. AHL22 and HDA15 fused with pSITE-nEYFP-N1 and pSITE-cEYFP-C1 vectors, respectively, were cotransformed into N. benthamiana. H3.3 RFP was used to indicate the nuclei. Scale bar = 50 um. d Quantification of the nuclei with both YFP and RFP signal from different pairwise of BiFC experiments. n = 120. Fisher’s exact test, **P < 0.01, ***P < 0.001. e FRET-FLIM analysis of AHL22 and HDA15 interaction in vivo. From top to bottom, representative images from nuclear signal, expressed by various GFP and/or RFP fused proteins in epidermal N. benthamiana (tobacco) cells, captured between wavelengths 540-545 nm, 570-670 nm and the corresponding donor fluorescence lifetime calibrated to the adjacent color bar. The respective construct combinations are indicated. Scale bar = 10 um. f GFP donor fluorescence lifetime quantifications of indicated construct combinations are presented in a min to max box and whiskers plot showing the median (line in the middle of the box), mean (+ in the box), min and max (whiskers), individual points (gray circle) (n > =18). Significance was calculated using a Welch’s ANOVA test combined with Dunnett’s T3 multiple comparisons test. P-values are represented as ****(p < 0.0001), ***(p < 0.001), **(p < 0.01). The experiment was performed in two independent infiltrations and the exact number of measurements was indicated per construct combination.
