Extended Data Fig. 3: Regulation of ERECTA protein abundance and specific pathways involved.

(a) RT-qPCR analysis of ERECTA in wild type and pub30 pub31 plants. Error bars represent SD (n = 3). Two tailed paired Student’s t-test was performed (p = 0.7073). ns, not significant. (b) ERECTA protein accumulation in wildtype, and pub30 pub31, in the absence and presence of the vacuolar ATPase inhibitor Concanamycin A (Con A). Total proteins were isolated from 7-d-old seedlings and probed by anti-FLAG antibody. The protein inputs were equilibrated using anti-actin antibody. (c) Quantification of ERECTA abundance (ERECTA/Actin) in the absence and presence of Tyr A23. Error bars represent SD (n = 3). The asterisks indicate statistical significance by using two-tailed paired Student’s t-test (p = 0.0067). (d) Quantification of ERECTA abundance (ERECTA/Actin) in the absence and presence of Con A. Error bars represent SD (n = 3). The asterisks indicate statistical significance by using two-tailed paired Student’s t-test (p = 0.0102). (e) The residues (W97 in PUB30 and W93 in PUB31) are essential for their autoubiquitination, respectively. The in vitro autoubiquitination assays were performed using GST-PUB30/31 wild type or mutants as the E3 ligases. (f) ERECTA protein accumulation in wild type, and pub30 pub31, in the absence and presence of the EPF2 and inactive EPF2. Total proteins were isolated from 7-d-old seedlings and probed by anti-FLAG antibody. The protein inputs were equilibrated using anti-actin antibody. (g) ERECTA protein accumulation in wild type, and pub30 pub31, in the absence and presence of the EPFL6 and inactive EPFL6. Total proteins were isolated from 7-d-old seedlings and probed by anti-FLAG antibody. The protein inputs were equilibrated using anti-actin antibody.