Extended Data Fig. 6: Full-length EIN2 is translocated to the nucleus in response to inhibition of TOR.
From: The TOR–EIN2 axis mediates nuclear signalling to modulate plant growth
a, b, EIN2–GFP and GFP–EIN2 complement the response of ein2-5 seedlings to treatment with Torin2 and ethylene (C2H4). Seedlings were germinated on solid half-strength MS medium with or without Torin2 (1 μM) or ethylene (100 ppm) in the dark for 4 days. Representative images (a) and hypocotyl length (b). Scale bar, 1 mm. Data were analysed from 22 seedlings for each genotype and treatment from 3 experiments, and are expressed as mean ± s.d. c, Different nucleus accumulation patterns of EIN2(WT)–GFP and GFP–EIN2(WT); see quantification data in Fig. 2g. Scale bars, 500 μm (left); 10 μm (right). d, EIN2 accumulates in the nucleus after inhibition of TOR. Arabidopsis mesophyll protoplasts transiently co-transfected with mCherry–EIN2–GFP and CFP–HDEL (an ER marker) were treated with or without rapamycin (10 μM) or Torin2 (1 μM) for 3 h. Scale bars, 5 μm. Experiments were repeated three times with similar results. e, f, Full-length EIN2 is translocated to the nucleus in response to inhibition of TOR. Four-day-etiolated pEIN2::EIN2WT-3XHA seedlings were treated with or without Torin2 (1 μM) for the indicated times, then total and nuclear fractions of hypocotyl protein were prepared and subjected to western blot (e). Tubulin (total protein loading control), calnexin homologue 1 or 2 (CNX1/2, ER marker) and histone H3 (nuclear marker) were used as controls to monitor the loading and purity of the total and nuclear fraction. f, Quantification of e. Data are mean ± s.d., n = 3 biological repeats. For b, f, statistical significance was determined by one-way ANOVA with Tukey’s post hoc test; n.s, not significant; P values are indicated. For gel source data, see Supplementary Fig. 1.
