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. 2019 Mar 19;180(2):1081–1100. doi: 10.1104/pp.18.01371

Figure 3.

Figure 3.

Glc regulates HLP1 transcription through TOR signaling. A, RT-qPCR expression of HLP1 in Col-0 and TOR overexpression lines (G166 and G548). Five-d–old Arabidopsis plants were first starved with 0% Glc for 24 h followed by Glc treatment (3% Glc for 3 h). Data shown are average of two biological replicates. Experiments were repeated two times with similar results. B, Effect of TOR kinase inhibitors on HLP1 expression. Five-d–old light-grown Arabidopsis seedlings were starved without Glc-containing MS media for 24 h under dark-grown conditions. After starvation, seedlings were treated with TOR inhibitors (Torin 1 and AZD-8055; 10 μM for 4 h in presence of 3% Glc). Data shown are average of two biological replicates. Experiments were repeated two times with similar results. C, Effect of glycolysis and mitochondrial inhibitors on HLP1 expression. Five-d–old light-grown Arabidopsis seedlings were starved in MS media without Glc for 24 h under dark-grown conditions. After starvation, seedlings were treated with glycolysis inhibitor (2-deoxy-Glc, 3%), mitochondrial electron transport inhibitor (antimycin A, 5 µM) and mitochondrial uncoupler (2,4-dinitrophenol 50 µM) for 4 h in presence of 3% Glc. Data shown are average of two biological replicates. Experiments were repeated two times with similar results. D, qPCR showing enrichment of HLP1 promoter fragments in all three regions after ChIP assays with or without Glc treatment (5 mM for 4 h). Protoplasts were harvested from 4-week–old Col-0 plants and subjected to Glc treatment. Fold enrichment of promoter fragments were calculated by comparing samples treated without or with E2Fa-specific antibody. Data shown are the average of four biological replicates. Experiments were repeated four times with similar results. ACT2 was taken as a control for the ChIP assay. E, RT-qPCR PCR showing expression of HLP1 in Col-0 and e2fa-1 mutant under HS. Arabidopsis Col-0 and e2fa-1 seedlings were grown in 1/2-strength MS medium containing 1% Suc for 5 d and then subjected to HS. HS was applied at 37°C for 3 h and the seedlings were harvested immediately. Data shown are the average of two biological replicates. Experiments were repeated two times with similar results. Ctrl, control. Error bars = sd (Student’s t test, P < 0.05; *control versus treatment; **wild type versus overexpression or mutant/3% Glc versus 3% Glc + inhibitors).