Fig 1: TRIM47 upregulates the expression of CCL15 and CCR1 via interacting with SMAD4 and enhancing ubiquitylation and degradation of SMAD4. a and b Western blot was performed in HCT116 and HT29 cells transfected with control siRNA and TRIM47 siRNAs (n = 3). c Western blot was performed in SW480 cells transfected with vector plasmids and TRIM47 overexpression plasmids (n = 3). d Western blot was performed in SW480 cells transfected with TRIM47 overexpression plasmids and treated with DMSO or MG132 (n = 3). e Western blot was performed in HCT116 cells transfected with TRIM47 siRNAs and treated with DMSO or MG132 (n = 3). f The amount of ubiquitin that co-immunoprecipitated with SMAD4 in SW480 cells transfected with TRIM47 overexpression plasmids. Western blot data of TRIM47, ubiquitin and GAPDH from 20% input (left). Anti-ubiquitin and anti-SMAD4 antibody were used for western blot to determine the ubiquitination level of SMAD4 (right) (n = 3). g The amount of ubiquitin that co-immunoprecipitated with SMAD4 in HCT116 cells transfected with TRIM47 siRNAs (n = 3). h The amount of ubiquitin that co-immunoprecipitated with SMAD4 in SW480 cells transfected with TRIM47 overexpression plasmids and the △RING domain plasmids (n = 3). RING, RING-finger domain; PR, proline-rich region; B-box, B-box-type zinc finger; CC, coiled coil region; SPRY, C-terminal SplA/ryanodine receptor domain; Flag-, Flag-tagged; △RING, deletion of the RING domain. i Co-immunoprecipitation detected the interaction of TRIM47 and SMAD4 in SW480 and HCT116 cells. The specific immunoprecipitation of TRIM47 and SMAD4 was confirmed by western blot (n = 3). j Immunofluorescence revealed that TRIM47 is co-localized with SMAD4 (n = 3)
Fig 2: TRIM47 overexpression promotes 5′-fluorouracil resistance of colorectal cancer cells in vitro and in vivo. a and b Cell proliferation assay was performed in SW480 cells transfected with vector plasmids, TRIM47 overexpression plasmids and SMAD4 overexpression plasmids (n = 3, non-parametric Mann-Whitney test, P < 0.05). c Cell transwell invasion assay was performed in SW480 cells transfected with vector plasmids, TRIM47 overexpression plasmids and SMAD4 overexpression plasmids (n = 3, non-parametric Mann-Whitney test, P < 0.05). d Cell proliferation assay was performed in SW480 cells treated with increasing concentrations of 5-FU. (n = 3, non-parametric Mann-Whitney test, P < 0.05). e Cell proliferation assay was performed in SW480 cells transfected with vector plasmids, TRIM47 overexpression plasmids and 5-FU (n = 3, non-parametric Mann-Whitney test, P < 0.05). f Representative images of tumors in nude mice bearing colorectal cancer cells treated with control adenovirus, TRIM47 overexpression adenovirus, 5-FU, TRIM47 overexpression adenovirus & 5-FU (n = 8). g Tumor volume was measured after different treatments in nude mice (n = 8, non-parametric Mann-Whitney test, P < 0.05). h Tumor weight was measured in nude mice after different treatments (n = 8, non-parametric Mann-Whitney test, P < 0.05). i A schematic model of TRIM47 function in colorectal carcinogenesis. TRIM47 interacts with SMAD4 and promotes the ubiquitination and degradation of SMAD4, impairing the suppression of CCL15 and CCR1, ultimately promoting colorectal cancer cells proliferation and metastasis
Supplier Page from R&D Systems, a Bio-Techne Brand for CCL15/MIP-1 delta 68 aa Protein
Available conjugates: Sizes Available: 25 ug