(A-B) Western blots showing EIN3 protein levels in the 4-day old seedlings grown in the dark (A) or white light (B). Seedlings over-expressing EIN3-Myc in ein3eil1 (E3Mee) and cop1ein3eil1 (E3Meec-4) backgrounds were grown on 1/2MS medium with or without 2.5mm soil (50-70 mesh Sand) covering. Bottom panels show the quantification analysis of the three biological replicates of EIN3-Myc protein levels after normalizing to Actin. The protein levels of EIN3-Myc/ein3eil1 without soil covering was set as 1. Mean ±s.d., n=3.
(C-D) Western blots showing the EIN3-Myc protein levels in ein3eil1 (C) or cop1ein3eil1 (D) background seedlings upon mechanical stresses. Seedlings were grown on 1/2MS medium for 4 days in the dark, then were pressed by a glass plate with the pressure of about 150Pa for the indicated periods of time. Bottom panels show the quantification analysis of the three biological replicates of EIN3-Myc protein levels after normalizing to Actin. The protein level of EIN3-Myc/ein3eil1 without mechanical stress was set as 1. Mean ±s.d., n=3.
(E) Light fluence in the soil increases as seedlings grow towards the soil surface. The light fluence was measured under the water-saturated soils with indicated particle sizes and depths.
(F) Western blots showing the EIN3 and HY5 protein levels in ein3eil1 and cop1ein3eil1 seedlings grown under indicated light fluences (μmolm2s-1) for 4 days. Dark-grown 4 day-old Col-0 was used as a negative control. Bottom panel shows the quantification analysis of the three biological replicates of EIN3-Myc protein levels after normalizing to Actin. The protein level of EIN3-Myc/ein3eil1 in the dark was set as 1. Mean ±s.d., n=3.