Fig. 5: SlCOL1 is directly activated by MIB2.
From: Genetic control of thermomorphogenesis in tomato inflorescences
a Chromatin binding profiles of MIB2, and ChIP-qPCR analysis of bound regions at the SlCOL1 promoter by resampling. The short black lines labeled P1, P2, P3, and P4 represent the regions used for ChIP-qPCR. IgG was used as a negative control. The DNA fragment of the ACTIN (Solyc03g078400) 3’ intergenic region was used as an internal control. b Electrophoretic mobility shift assay (EMSA) of recombinant MBP-MIB2 protein with a biotin-labeled DNA fragment containing the G-box motif of the SlCOL1 promoter. The arrow indicates the MIB2–DNA complex. Three independent experiments were performed. c Representative dual-luciferase reporter assay in N. benthamiana co-infiltrated with pro35S:MIB2-FLAG or pro35S:FLAG and proSlCOL1:LUC-pro35S:REN. Leaves co-infiltrated with proSlCOL1: LUC-pro35S: REN and pro35S:FLAG were used as a control. Renilla luciferase (REN) was used as an internal control. Data were compared by two-tailed Student’s t-test, **, p < 0.01. Values are means ± SEM. N = biologically independent samples. d, e Relative SlCOL1 expression in meristematic tissues of NIL-MIB2ST082 and mib2cr plants (d) and MM and proMIB2:MIB2-YFP-HA complementation plants (e). UBIQUITIN3 (Solyc01g056940) was used as an internal control. f ChIP-qPCR analysis showing the enrichment of MIB2 at the SlCOL1 promoter after high temperature (35°C) treatment. Meristematic tissues were collected from proMIB2:MIB2-YFP-HA transgenic plants and analyzed at the indicated time points. P1, P2, P3 and P4 represent the regions used for ChIP-qPCR. IgG was used as a negative control. g Relative SlCOL1 expression in the meristematic tissues of NIL-MIB2ST082 and mib2cr plants after 8 hours of 35°C treatments. Seedlings were grown to the 4-leaf stage at 22 °C under moderate sunlight, transferred to 22 °C in full sunlight and grown for 5 days, and transferred to 35 °C and grown for 8 hours in full sunlight. h Inflorescence phenotypes (left) and inflorescence branch number (right) of NIL-mib2MM and SlCOL1 overexpression transgenic plants (SlCOL1-OEs) under HT conditions. N = inflorescence number. Scale bars, 3 cm. Data in (a, d, e, g) were compared by two-tailed Student’s t-test, **, p < 0.01; n.s. no significant difference (p > 0.05). Values are means ± SEM (n = 4 biologically independent replicates). Different letters in (f) indicate a significant difference (p  <  0.05) based on the one-way ANOVA followed by Tukey’s multiple comparisons test. Values are means ± SEM (n = 4 biologically independent replicates). Source data are provided as a Source Data file.
