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. 2009 Feb 16;4(2):e4431. doi: 10.1371/journal.pone.0004431

Figure 1. Pull-down assays with GST-YopH D356A.

Figure 1

A, HEK293 cells expressing different proteins, either untreated (control) or treated with pervanadate (PV) to induce tyrosine phosphorylation of the proteins expressed, were lysed and probed for interaction with GST-YopH D346A or GST (5 µg each) as a negative control in pull-down assays. The specific interaction of those proteins with GST-YopH D356A was detected by Western blot with specific antibodies for Lck or Vav, and with anti-HA antibody for other proteins. B, As in A, HEK293 cells, expressing the same proteins and treated with PV, were lysed and probed for interaction with two different amounts of GST-YopH D346A (5 and 2 µg) or GST (5 µg each) as a negative control in pull-down assays. GST and GST-YopH D346A fusion protein used in these assays are shown at the lower panel from one representative blot. TL denotes total lysates of the transfected cells and corresponds to a 10% of the amount used for each pull-down assay. Assays were done independently for each protein.