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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 2.

FIGURE 2

SlNRC4a and LeEIX2 associate in planta when expressed in Nicotiana benthamiana. (a) Plants were transiently cotransformed with LeEIX2-GFP or LeEIX2 and SlNRC4a-mCherry. Leaves were harvested and treated with either EIX or water (mock) at the petiole for 15 min. Triton X-100 soluble membrane (TSM) protein fractions were immunoprecipitated (IPed) using GFP affinity beads. Input IP samples were subjected to sodium dodecyl sulphate-polyacrylamide gel electrophoresis and immunoblot analyses. Membranes were probed with anti-GFP antibodies to detect LeEIX2-GFP and anti-mCherry antibodies to detect SlNRC4a-mCherry. (b) SlNRC4a-mCherry signal intensity was determined using Coloc2 from FIJI-ImageJ, by dividing co-IP intensity by input intensity. SlNRC4a-mCherry signal intensity, without EIX treatment, was defined as 100%. Error bars represent the average ± SD of three independent experiments. Asterisks indicate significant differences (t test, *P < 0.05, **P < 0.005, ***P < 0.001)