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. 2011 Jan 20;286(11):9373–9381. doi: 10.1074/jbc.M110.207720

FIGURE 1.

FIGURE 1.

TIP30 interacts with Rab5a, ACSL4, and Endo B1. A, immunoblot confirmed the association of Rab5a, ACSL4, and Endo B1 with TIP30. The immunoprecipitates were resolved on SDS-PAGE and analyzed by immunoblotting (IB) with the indicated antibodies. The two polypeptides that reacted with anti-HA antibodies might result from posttranslational modifications on TIP30. Input was 10% (equals 70 μg of protein) of the volume of lysates used in the immunoprecipitation (IP) assays. The EGFR blot serves as a negative control. B, reciprocal co-immunoprecipitation. Whole cell extracts made from cells expressing HA-Rab5a, ACSL4-HA, Endo B1-HA, or empty vector were subjected to co-immunoprecipitation with α-HA-agarose beads. The immunoprecipitates were subjected to immunoblot analysis with the indicated antibodies. Input was 10% of the volume of lysates used in the immunoprecipitation assays. C, bimolecular fluorescence complementation analysis was performed on 293T cells by co-expressing TIP30-VC155 with VN173, VN173-Rab5a, ACSL4-VN173, or Endo B1-VN173. Green indicates fluorescent signal from Venus, a GFP variant. Bottom panels show overlays of differential interference contrast and representative confocal microscope images. Scale bars, 10 μm.