Fig. 4: SlARF2a–TSP4a and TSP4b directly regulate the transcriptions of TSP4b and TSP4a. | Nature Communications

Fig. 4: SlARF2a–TSP4a and TSP4b directly regulate the transcriptions of TSP4b and TSP4a.

From: A feedback loop at the THERMOSENSITIVE PARTHENOCARPY 4 locus controls tomato fruit set under heat stress

Fig. 4

a ChIP–qPCR assay of TSP4b–YFP–HA enrichment at the TSP4a promoter. Schematic of promoter regions and primer-binding sites used for ChIP–qPCR (P1–6). P1 region was used as a negative control and Input was mock-IP negative control without the antibody. b Representative dual-luciferase reporter assay in N. benthamiana co-infiltrated with 35Spro:FLAG or 35Spro:TSP4b–FLAG and TSP4apro:LUC-35Spro:REN. Leaves co-infiltrated with TSP4apro:LUC-35Spro:REN and 35Spro:FLAG were used as control (n = 8 biologically independent replicates). c Yeast two-hybrid of the protein interaction between TSP4a and SlARF2a. Yeast cells were grown on -Leu/-Trp (-LT) plates and -Leu/-Trp/-Ade/-His (-LTHA) plates (containing 30 mM 3-AT) for 3 d. d BiFC visualization of the interaction between TSP4a and SlARF2a in the nuclei of N. benthamiana epidermal cells, Bars = 50 μm. Three independent experiments were conducted with similar results. e Fruits of Moneymaker and 35Spro:SlARF2aTr–YFP–HA over-expression lines. Scale bar = 2 cm. SlARF2a expression in –2 DPA ovaries and seedless fruit rate from Moneymaker and 35Spro:SlARF2aTr–YFP–HA under heat stress (n = 5 plants). f ChIP–qPCR assay of SlARF2aTr–YFP–HA enrichment at the TSP4b 3’ UTR. Schematics of promoter regions and primer-binding sites used for ChIP–qPCR (P1–7). P1 region was used as a negative control and Input was mock-IP negative control without the antibody. g TSP4b expression in –2 DPA ovaries isolated from Moneymaker and 35Spro:SlARF2aTr–YFP–HA plants under heat stress in the greenhouse. h Representative dual-luciferase reporter assay in N. benthamiana co-infiltrated with 35Spro:FLAG, 35Spro:TSP4a–FLAG, 35Spro:SlARF2a–FLAG, 35Spro:SlARF2aTr–FLAG and 35Sminpro:LUC-TSP4butr-35Spro:REN. Leaves co-infiltrated with 35Sminpro:LUC-TSP4butr-35Spro:REN and 35Spro:FLAG were used as a control (n = 10 biologically independent replicates). Data in (a, e–g) are the mean ± S.D. of n = 3 biologically independent replicates, qPCR and ChIP-qPCR were normalized to UBIQ3 and ACTIN, respectively. Data in (a, b, f) were determined by two-tailed Student’s t-test. Data in (e, g, h) were compared by one-way ANOVA with multiple comparisons and Tukey post-hoc test (α = 0.05). **P < 0.01, n.s. non-significant (P > 0.05). Source data are provided as a Source Data file.

Back to article page