Analysis of exosome secretion in wild-type and RHBDD1-mt HCT116 cells.

A. A schematic diagram of the targeting motif of RHBDD1 knock-in. B. Quantitative analysis of total exosomes secreted from 2×10⁷ HCT116 cells for 16 h. C. Immunodetection of Tsg101 and Tf-R contents in 4 µg of exosomal proteins from wild-type and RHBDD1-mt HCT116 cells. TSAP6 expression was normalized with GAPDH and shown below (** P<0.01). D. Real-time analysis of the relative mRNA levels of RHBDD1, TSAP6 and Bik in wild-type and RHBDD1-mt HCT116 cells. E. Western blot analysis of Tsg101 and Tf-R proteins on fractions collected from 40 µg of total exosome samples from wt and RHBDD1-mt HCT116 cells processed on a 0.25–2.5 M sucrose gradient.