Fig. 8.

CRC cell‐derived exosomal STAT3 promotes Cyr61 transcription. (A) Western blot analysis of p‐STAT3 and STAT3 expression in ADSCs‐NC and ADSCs‐CRC. (B) qRT‐PCR analysis of Cyr61 mRNA levels in ADSCs treated with STAT3 inhibitors. n = 3. (C) Luciferase activity of the Cyr61 promoter in ADSCs transfected with STAT3. n = 3. (D) Luciferase (luc) reporter assays for ADSCs transfected with a series of reporter vectors containing sequential deletion of the Cyr61 promotor. n = 3. (E) Potential STAT3‐binding site in the Cyr61 promoter predicted in the JASPAR database. Luciferase reporter assays for ADSCs transfected with reporter plasmids containing point mutation at the STAT3‐binding site. n = 3. (F, G) qRT‐PCR analysis of the product of chromatin immunoprecipitation assays for the binding of STAT3 to the crucial STAT3‐binding site in the Cyr61 promoter. (H) Western blot analysis of p‐STAT3 and STAT3 expression changes in ADSCs cocultured with HC8 cells. (I) qRT‐PCR analysis of Cyr61 mRNA levels in ADSCs cocultured with HC8 cells. n = 3. (J) Western blot analysis of p‐STAT3 and STAT3 in normal colonic cell and CRC cell‐derived exosomes. (K) Western blot analysis of p‐STAT3 and STAT3 expression in ADSCs cocultured with CRC cell‐derived exosomes with or without p‐STAT3 and STAT3 knockdown. (L) qRT‐PCR analysis of Cyr61 mRNA levels in ADSCs cocultured with CRC cell‐derived exosomes with or without p‐STAT3 and STAT3 knockdown. n = 3. (M) Schematic model of the role of Cyr61 in CRC tumor progression. Values are represented as mean ± SD. CO, cocultured; EXO, exosomes. NS, no significant, **P < 0.01, ***P < 0.001, by two‐tailed Student's t‐test (C, E, F, I, L) and one‐way ANOVA (B, D).