Extended Data Fig. 10: Complementary assay of NSP1s and NSP13KR mutant, monoubiqutination specificity assay of NSP1a by BTSs. | Nature Plants

Extended Data Fig. 10: Complementary assay of NSP1s and NSP13KR mutant, monoubiqutination specificity assay of NSP1a by BTSs.

From: The BRUTUS iron sensor and E3 ligase facilitates soybean root nodulation by monoubiquitination of NSP1

Extended Data Fig. 10: Complementary assay of NSP1s and NSP13KR mutant, monoubiqutination specificity assay of NSP1a by BTSs.The alternative text for this image may have been generated using AI.

a, Nodulation phenotype of transgenic soybean hairy roots in wild-type Wm82 and nsp1a-1 heterologously expressing UBIpro:MtNSP1WT–4×MYC, UBIpro:LjNSP1WT–4×MYC, UBIpro:MtNSP13KR–4×MYC or UBIpro:LjNSP13KR–4×MYC constructs. Nodules were harvested at 28 DAI. Scar bars = 2 cm. The experiments were repeated for three times with similar results. b, In vivo ubiquitination assay of NSP1a by AtBTS. Proteins extracted from tobacco leaves transiently transformed with (+) or without (–) 35Spro:NSP1a–3×FLAG, 35Spro:UBQ–6×MYC, UBIpro:BTSa–3×HA, UBIpro:AtBTSa–3×HA and empty vectors. Anti-MYC agarose was used for immunoprecipitation. The experiment was repeated for three times with similar results. c, Co-IP assay to test the interaction of AtBTSa–3×HA and NSP1a–3×FLAG or NSP2a–3×FLAG in N. benthamiana leaves. Protein was immunoprecipitated with HA antibody-bound agarose beads, immunoprecipitated proteins were analyzed using anti-FLAG and anti-HA antibodies. The experiments were repeated for three times with similar results.

Source data

Back to article page