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Fig. 3 | BMC Plant Biology

Fig. 3

From: The ARGOS gene family functions in a negative feedback loop to desensitize plants to ethylene

Fig. 3

ARGOS and ARL are membrane-associated proteins localized to the ER. a Membrane association of ARGOS-GFP and ARL-GFP. Membrane and soluble fractions were isolated from green seedlings following 4-h treatment in the absence or presence of 10 μL L−1 ethylene. Immunoblot analysis was performed with anti-GFP antibody to detect the ARGOS-GFP and ARL-GFP. Immunological detection of the ethylene receptor ETR1 and ACC-oxidase served as markers for the membrane and the soluble fractions, respectively. b Strong association of ARGOS and ARL with membranes. Microsomal membranes were treated with 0.5 M NaCl or 0.5% (w/v) lysophosphatidylcholine (LPC). The different lanes represent the total membranes prior to centrifugation (T), and from the soluble (S) and (P) fractions after centrifugation. The relative amounts of ARGOS-GFP, ARL-GFP, and ETR1 were determined by immunoblot analysis, the membrane protein ETR1 serving as an internal control for solubilization. c ARGOS and ARL localize to the ER. Protoplasts were transfected with either ARGOS-GFP or ARL-GFP (green), along with the ER-marker BiP-RFP (magenta), and visualized by confocal microscopy. Regions of overlap are indicated by white on the merged image. DIC images of protoplasts are also shown. Focused region is on the cortical ER underlying the plasma membrane. Scale bars = 10 μm

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