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1.
Figure 2

Figure 2. Group VII ERF transcription factors are substrates for the N-end rule pathway in vitro and in vivo. From: Homeostatic response to hypoxia is regulated by the N-end rule pathway in plants.

a. Western blot analysis of in vitro stability of HA-tagged WT and C2A variants of Arabidopsis Group VII ERFs in the absence or presence of MG132, N-end rule pathway competitive dipeptide (Arg-β-Ala) or noncompetitive dipeptide (Ala-Ala).
b. In vitro stability of WT and C2A VRN2-HA and MAF5-HA.
c. In vitro stability of HA-tagged rice ERFs.
d. In vivo protein stability and RNA expression levels of WT and C2A variants of HRE1-HA and HRE2-HA ectopically expressed in Arabidopsis, shown for two independent transformed lines (1 and 2).

Daniel J. Gibbs, et al. Nature. ;479(7373):415-418.
2.
Figure 1

Figure 1. N-end rule mutants ectopically accumulate anaerobic response mRNAs and are more tolerant to hypoxia. From: Homeostatic response to hypoxia is regulated by the N-end rule pathway in plants.

a. Expression data for differentially expressed genes comparing WT (Col-0) and mutants under air or hypoxia (2 h –O2).
b. mRNAs upregulated in mutants overlap with 49 mRNAs induced across cell types by hypoxia in WT seedlings15.
c. Spatial visualization of ADH1 promoter activity. Scale bars 100 μm.
d. Germination under reduced oxygen availability.
e. Seedlings after 12 h of hypoxia and 3 d recovery. Scale bar 0.6 cm.
f. N-end rule pathway mutants are less sensitive to hypoxia stress. Data are mean of replicate experiments ± SD; * = P < 0.05, ** = P < 0.01.

Daniel J. Gibbs, et al. Nature. ;479(7373):415-418.
3.
Figure 3

Figure 3. HRE proteins are stabilized under low oxygen and confer hypoxia tolerance. From: Homeostatic response to hypoxia is regulated by the N-end rule pathway in plants.

a. In vivo stability of WT and C2A HRE1-HA, HRE2-HA (α-HA) or S6 (Ribosomal protein S6) control (α-S6): no stress (NS), 2 h hypoxia (HS), following 1 h recovery from stress (R).
b. Seedlings expressing WT or C2A HRE1-HA and HRE2-HA after 12 h hypoxic stress and 3 d of recovery. Scale bar = 0.6 cm.
c. Seedling survival for WT or C2A HRE1-HA and HRE2-HA after 9 h or 12 h hypoxic stress. Data are mean of replicate experiments ± SD; * = P < 0.05, ** = P < 0.01.
d. Germination under reduced oxygen availability.
e. Model explaining N-end rule pathway mediated oxygen-dependent turnover of Group VII ERFs in Arabidopsis.

Daniel J. Gibbs, et al. Nature. ;479(7373):415-418.

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