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. 2003 Aug;132(4):2023–2033. doi: 10.1104/pp.103.020438

Figure 4.

Figure 4.

NHL3-HA in transgenic Arabidopsis is a glycosylated and membrane-associated protein complex. A, Microsomal fractions of tobacco leaves transiently expressing NHL3-HA were incubated for 30 min with buffer (M-) or with PNGase F (M+) and were subjected to western-blot analysis with the anti-HA antibody. B, Microsomal fractions isolated from leaves of DEX-treated transgenic NHL3-HA Arabidopsis plants were treated and analyzed as described in A. C, Higher mobility complexes (arrows) were seen when the protein extracts were treated with the indicated amount of chemical crosslinker, 3,3′-dithiobis(sulfosuccinimidylpropionate) (DTSSP) before western blotting. D, Microsomal fractions isolated from leaves of DEX-treated transgenic NHL3-HA Arabidopsis plants were treated with the indicated substances for 1 h at 4°C. Membranes were then pelleted by ultracentrifugation and the resulting soluble (S) and membrane proteins (M) were analyzed by western blotting.