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. Author manuscript; available in PMC: 2020 Jun 2.
Published in final edited form as: Plant Cell Environ. 2018 Jul 3;41(10):2313–2327. doi: 10.1111/pce.13347

FIGURE 4.

FIGURE 4

(a) SlNRC4a and FLS2 associate in planta when expressed in Nicotiana benthamiana. Plants were transiently cotransformed with pAtFLS2::AtFLS2-GFP or NOS::GFP and 35S::SlNRC4a-2xHA using Agrobacterium infiltration. Forty-eight hours after infiltration, leaves were harvested and treated with either 1 mM of flg22 or water (mock) at the petiole for 15 min; 0.5 g of leaves per treatment was used for protein extraction. Triton X-100 soluble membrane protein fractions were subjected to anti-GFP IP. Input and IPed proteins were subjected to sodium dodecyl sulphate-polyacrylamide gel electrophoresis and immunoblot analyses with anti-GFP and anti-HA. (b) Effect of overexpression of SlNRC4a in N. benthamiana on flagellin-mediated ROS burst. Leaf disks of transiently transformed N. benthamiana leaves with SlNRC4a-GFP or control (free GFP) as indicated (24 hr after transformation) were floated on water for ~24 hr and then replaced with luminescence solution containing 1 μM of flg22. Luminescence was immediately measured to track the ROS burst. Average ± SEM values of three independent experiments, n = 12 each. Asterisks indicate significant differences with control treatment (two-way analysis of variance, *P < 0.05, **P < 0.01, ***P < 0.001)