Fig. 6

LAP-TGF-β1-HSP90A interaction directs transport from Golgi to endosomes. (A) Post-transfection LAP-TGF-β1 exosomal expression from 231 LuT3 cells transfected with designated siRNA (n = 3). (B) Exosomal LAP-TGF-β1 levels after 231 parental cell transfection with either His-vector or His-HSP90AA1 (n = 3). (C-E) Co-transfection of 231 parental cells with His-HSP90AA1 and HA-LAP-TGFB1, followed by cell purification using specific agarose beads and subsequent antibody-based analysis. (F) LAP-TGF-β1-containing exosomes after 231 LuT3 cell transfection with si HSP90AA1 or si NC (n = 3). (G) HA-LAP-TGFB1 and si HSP90AA1 or si NC were co-transfected into 231 LuT3 cells and purified using anti-HA agarose beads. Immunoprecipitated and immunoblotted cell lysates using the appropriate antibodies. (H-L) Specific co-transfections and their subsequent intracellular interactions visualized using color-coded labels (n = 5). Scale bars: 10 μm and 5 μm. Data shown as mean ± SD