Fig. 4.

TRIM6 interacted with TIS21 in CRC cells. a, pCMV-Tag2-TIRM6 or pCMV-Tag2 vector was transfected into 293 T cells, and 48 h later, cell lysates were prepared and subjected to immunoprecipitation (IP) experiments with anti-FLAG beads. After elusion with FLAG peptide, the immunoprecipitated protein complexes were resolved on SDS-PAGE, and stained with Coomassie Brilliant Blue. B, C, IP was carried out with TRIM6 antibody (TRIM6-Ab) /IgG (b) or TIS21 antibody (TIS21-Ab) /control IgG (c), and then western blotting was performed to analyze specific associations between TRIM6 and TIS21 in HCT-8 and HCT116 cells. D-E, GST pull-down assay. HCT-8 cells were lysed and incubated with GST, GST-tagged TRIM6 (d) and GST-tagged TIS21 (e) bound to glutathione beads, respectively. Proteins were detected as indicated. E, immunofluorescence staining of TRIM6 (Red) and TIS21 (Green) in HCT-8 and HCT116 cells. DAPI (blue) was used to label nuclei. Scale bar: 50 μm