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Figure 6 | BMC Plant Biology

Figure 6

From: The Arabidopsis apyrase AtAPY1 is localized in the Golgi instead of the extracellular space

Figure 6

Analysis of the solubility of AtAPY1-GFP. (A) The output of the TMHMM prediction program [69] is shown. The AtAPY1 aa sequence is represented along the x-axis with the N-terminal aa being the first residue. The probability for each aa to appear within a transmembrane domain (TM) is given. One TM is predicted which comprises the aa 21–43. (B) Microsomal membranes were prepared from transgenic plants containing 35S::AtAPY1-GFP. Equal amounts of membrane fractions were treated either with extraction buffer, 0.2% SDS, 2 M NaCl, 0.2 M Na2CO3 or 4 M urea. After a 30-min treatment, the proteins were subjected to centrifugation at 100,000 g. The pellet fractions (P) and the supernatants (S) were separated in a 8% SDS polyacrylamide gel, blotted on a nitrocellulose membrane and incubated with α-GFP antibodies. The molecular weights of the protein standard are given in kDa. The AtAPY1-GFP protein (80 kDa) and a second band at 27 kDa (arrow) were detected with α-mouse IgG coupled with horseradish peroxidase. Similar results were obtained in three experiments with independent protein extracts.

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