Figure 5
AtMMS21 mediates the SUMOylation of IAA17 regulating IAA17 degradation. A, IAA17 protein levels in UBQ:IAA17-GFP/WT and UBQ:IAA17-GFP/mms21-1. UBQ:IAA17-GFP/WT and UBQ:IAA17-GFP/mms21-1 were treated with 1/2 MS for 0, 10, and 20 min. The protein levels of IAA17-GFP in the seedlings were detected by immunoblotting analysis using a GFP antibody. The loading controls from Coomassie Blue staining are shown at the bottom. B, Quantification of the IAA17-GFP blot in (A). The quantification of IAA17-GFP in 0 min is set to 1. The data are means ± Sd from 2 biological independent experiments. C, IAA17 protein levels in UBQ:IAA17-GFP/WT and UBQ:IAA17-GFP/mms21-1 under NAA treatment. UBQ:IAA17-GFP/WT and UBQ:IAA17-GFP/mms21-1 were treated with 1/2 MS containing 100 µM NAA for 0, 10, and 20 min. The protein levels of IAA17-GFP in the seedlings were detected by immunoblotting analysis using a GFP antibody. The loading controls from Coomassie Blue staining are shown at the bottom. D, Quantification of the IAA17-GFP blot in (C). The quantification of IAA17-GFP in 0 min is set to 1. The data are means ± Sd from 2 biological independent experiments. *P < 0.05, ***P < 0.001, Student's t test. E, IAA17 protein levels in UBQ:IAA17-GFP/WT and UBQ:IAA17-GFP/mms21-1 under NAA and MG132 treatment. UBQ:IAA17-GFP/WT and UBQ:IAA17-GFP/mms21-1 were treated with 1/2 MS containing 100 µM NAA and 100 µM MG132 for 0, 10, and 20 min. The protein levels of IAA17-GFP in the seedlings were detected by immunoblotting analysis using a GFP antibody. The loading controls from Coomassie Blue staining are shown at the bottom. F, Quantification of the IAA17-GFP blot in (E). The quantification of IAA17-GFP in 0 min is set to 1. The data are means ± Sd from 2 biological independent experiments. G, IAA17-GFP signals in the roots of UBQ:IAA17-GFP/WT, UBQ:IAA17K41R-GFP/WT, and UBQ:IAA17-GFP/mms21-1 seedlings under NAA treatment. Five-day-old seedlings were treated with 100 µM NAA for 5 min. and the GFP signals were observed and collected via microscopy with the same settings. Bars, 50 μm. NAA, 1-Naphthylacetic acid.

AtMMS21 mediates the SUMOylation of IAA17 regulating IAA17 degradation. A, IAA17 protein levels in UBQ:IAA17-GFP/WT and UBQ:IAA17-GFP/mms21-1. UBQ:IAA17-GFP/WT and UBQ:IAA17-GFP/mms21-1 were treated with 1/2 MS for 0, 10, and 20 min. The protein levels of IAA17-GFP in the seedlings were detected by immunoblotting analysis using a GFP antibody. The loading controls from Coomassie Blue staining are shown at the bottom. B, Quantification of the IAA17-GFP blot in (A). The quantification of IAA17-GFP in 0 min is set to 1. The data are means ± Sd from 2 biological independent experiments. C, IAA17 protein levels in UBQ:IAA17-GFP/WT and UBQ:IAA17-GFP/mms21-1 under NAA treatment. UBQ:IAA17-GFP/WT and UBQ:IAA17-GFP/mms21-1 were treated with 1/2 MS containing 100 µM NAA for 0, 10, and 20 min. The protein levels of IAA17-GFP in the seedlings were detected by immunoblotting analysis using a GFP antibody. The loading controls from Coomassie Blue staining are shown at the bottom. D, Quantification of the IAA17-GFP blot in (C). The quantification of IAA17-GFP in 0 min is set to 1. The data are means ± Sd from 2 biological independent experiments. *P < 0.05, ***P < 0.001, Student's t test. E, IAA17 protein levels in UBQ:IAA17-GFP/WT and UBQ:IAA17-GFP/mms21-1 under NAA and MG132 treatment. UBQ:IAA17-GFP/WT and UBQ:IAA17-GFP/mms21-1 were treated with 1/2 MS containing 100 µM NAA and 100 µM MG132 for 0, 10, and 20 min. The protein levels of IAA17-GFP in the seedlings were detected by immunoblotting analysis using a GFP antibody. The loading controls from Coomassie Blue staining are shown at the bottom. F, Quantification of the IAA17-GFP blot in (E). The quantification of IAA17-GFP in 0 min is set to 1. The data are means ± Sd from 2 biological independent experiments. G, IAA17-GFP signals in the roots of UBQ:IAA17-GFP/WT, UBQ:IAA17K41R-GFP/WT, and UBQ:IAA17-GFP/mms21-1 seedlings under NAA treatment. Five-day-old seedlings were treated with 100 µM NAA for 5 min. and the GFP signals were observed and collected via microscopy with the same settings. Bars, 50 μm. NAA, 1-Naphthylacetic acid.

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