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. 2014 May 1;10(5):e1004102. doi: 10.1371/journal.ppat.1004102

Figure 4. Only Dlg1-I3 mediates E4-ORF1-induced PI3K activation whereas both Dlg1-I3 and Dlg1-I2 mediate PI3K protein elevation.

Figure 4

Left panel, The Dlg1 shRNA-expressing MCF10A line was transfected with expression plasmid GW1-E4-ORF1 (75 ng), GW1-HA-ΔNT-Dlg1-I3 (2 µg), or GW1-HA-ΔNT-Dlg1-I2 (4 µg) alone or in the indicated combinations. The total amount of DNA in each transfection was equalized to 7.575 µg using empty GW1 plasmid. At 48 h post-transfection, cells were serum starved for 1 h, and then extracts were prepared and analyzed in immunoblot assays using the indicated antibodies. The insoluble (I) pellet fraction (see Materials and Methods ) was immunoblotted with E4-ORF1 antibody. Right panel, This blot verifies Dlg1 protein depletion in the Dlg1 shRNA-expressing MCF10A line (+) used in the left panel by comparison to the control Dlg1 scrambled shRNA-expressing MCF10A line (−).