Research Paper Volume 12, Issue 21 pp 21809—21836

CCL5-dependent mast cell infiltration into the tumor microenvironment in clear cell renal cell carcinoma patients

Figure 7. High CCL5 expression and secretion correlates with mast cell infiltration in PBRM1MUTccRCC cells and patients. (A) Volcano plot shows fold changes in gene expression in control and PBRM1-overexpressing Caki-2 cells. The association of immune response with mutations in PBRM1, VHL, SETD2 and BAP1 genes is shown in black circles. (B) qRT-PCR analysis shows CCL5 mRNA expression in 786-O- and Caki-1-silenced PBRM1 cells. (C) ELISA assay results show CCL5 levels in the conditioned media of control and PBRM1-silenced 786-O and Caki-1 cells using the human CCL5 ELISA kit. (D) Transwell migration assay results show total numbers of migrating HMC-1 cells when co-cultured with conditioned media derived from control, PBRM1-silenced and PBRM1-silenced plus CCL5-silenced 786-O cells. The migrating MHC-1 cells were stained with crystal violet and counted. Note: All experiments were performed in triplicate and are presented as means±SD; statistical analysis was performed using Student’s t-test.