Figure 5.

ebs4 Can Suppress the bri1-5 Mutation.

(A) Four-week-old soil-grown plants of bri1-5 and ebs4-2 bri1-5.

(B) Two-week-old soil-grown plants of bri1-5 and ebs4-3 bri1-5.

(C) Five-day-old dark-grown seedlings of bri1-5 and ebs4-3 bri1-5.

(D) Seven-week-old soil-grown mature plants of bri1-5 and ebs4-3 bri1-5.

(E) Immunoblot analysis of BRI1 and bri1-5 with or without Endo H treatment in 3-week-old seedlings. Equal amounts of total proteins extracted in 1× SDS buffer from 3-week-old seedlings were treated with or without 1000 units of Endo H for 1 h at 37°C, separated by SDS-PAGE, and analyzed by immunoblotting with anti-BRI1 antibody. bri1-5^ER is the ER-localized form, while bri1-5^CT indicates bri1-5 carrying C-type N-glycans.

(F) Immunoblot analysis of the BES1 phosphorylation status in 3-week-old seedlings treated with or without 1 μM BL. Total proteins were extracted in 1× SDS buffer from 3-week-old seedlings treated with or without 1 μM BL for 1 h in liquid half-strength MS medium, separated by 10% SDS-PAGE, and analyzed by immunoblotting with anti-BES1 antibody. pBES1 is the phosphorylated form of BES1. In both (E) and (F), Coomassie blue staining of RbcS serves as the loading control. [See online article for color version of this figure.]