Fig. 1. CD147 is identified as a TFF3-binding protein and is indispensable for promoting cancer progression and activating downstream signaling by TFF3.
a Schematic representation illustrating the strategy of using His6-tagged TFF3 to capture interacting proteins from CRC tissues and of characterizing the interacting partner via LC-MS/MS. b Western blotting analyses of endogenous TFF3 co-IP with CD147 from CRC tissues. IgG was used as a control antibody. c Representative images of immunofluorescence staining of endogenous TFF3 and CD147 in CRC tissues. Scale bar, 50 μm. d Biacore diagram of human TFF3 protein bound to CD147ECD. The KD values were calculated by the Biacore T200 evaluation software. e Representative confocal images of HCT-8 and HCT-8 CD147KO cells incubated with Alexa Fluor™ 555-labeled TFF3. Scale bar, 10 μm. f Representative images of HCT-8 and HCT-8 CD147KO cells invading through matrigel-coated transwell inserts toward serum for 24 h. Cells were treated with 0.152 μM of TFF3 or BSA. Scale bar, 50 μm. The graph shows the average number of invaded cells per field. g Proliferation curve of HCT-8 and HCT-8 CD147KO cells determined by CCK-8 assays. Cells were treated with 0.152 μM of TFF3 or BSA. Significance relative to the WT+BSA group was determined by a two-tailed Student’s t-test. h Western blotting analyses of the indicated proteins in HCT-8 or HCT-8 CD147KO cells treated with 0.152 μM of TFF3. Graphs show semi-quantitative analysis of relative p-STAT3 and p-ERK1/2 expression. The p-values in f–h were determined by using two-tailed Student’s t-test