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INSIG1 and TRC8 did not interact with Pr55Gag directly. In 293T cells, TRC8-EGFP was co-transfected with HA-INSIG1 or Gag-pol, and INSIG1-EGFP was co-transfected with Gag-pol. Chloroquine was added to inhibit the degradation of Gag. Cell lysates were immunoprecipitated with GFP-Trap Sepharose and analyzed by Western blotting. INSIG1 was co-immunoprecipitated with TRC8 efficiently (A), but Gag did not co-immunoprecipitate with either INSIG1 (B) or TRC8 (C).
