Furthermore, the growth-related signaling pathways were detected by Western blot.  Furthermore, Western blot analysis showed that the expression of non-phosphorylated (activated) β-catenin and its target genes cyclin D1 as well as c-myc were distinctly increased in co-cultured SW480 cells, which could be reversed by anti-LL-37 antibodies. 
THERM
Methods: Colorectal cancer cells SW480 or HCT116 were co-cultured with human macrophages using Transwell(®) maxicell inserts to mimic the tumor microenvironment. 
Transwell(®) maxicell inserts
The effect of macrophages on the proliferation of colorectal cancer cells was detected by Bromodeoxyuridine and enzyme-linked immunosorbent assay (BrdU-ELISA). 
Bromodeoxyuridine and enzyme-linked immunosorbent assay (BrdU-ELISA)
The expression of LL-37 mRNA and protein in macrophages and colorectal cancer cells was evaluated by reverse transcription-real-time quantitative PCR (RT-qPCR) and Western blot. 
reverse transcription-real-time quantitative PCR (RT-qPCR)
The expression of LL-37 mRNA and protein in macrophages and colorectal cancer cells was evaluated by reverse transcription-real-time quantitative PCR (RT-qPCR) and Western blot.  Furthermore, the growth-related signaling pathways were detected by Western blot.  Consistently the protein expression of LL-37 in macrophages was significantly increased by Western blot, while it did not change in SW480.  Furthermore, Western blot analysis showed that the expression of non-phosphorylated (activated) β-catenin and its target genes cyclin D1 as well as c-myc were distinctly increased in co-cultured SW480 cells, which could be reversed by anti-LL-37 antibodies. 
Western blot