Figure 5
Conditioned medium (CM) from BRAFV600E cells increases the production of CXCR4 and SDF-1 by human dermal fibroblasts. Conditioned medium was collected from Bowes cells (clones 7, 8, and 21) that had been incubated for 48 h in serum-free medium either with or without 3 μ M PLX4032. This CM was placed on human dermal fibroblasts (‘Fibroblasts’) and allowed to incubate on these cells for 24 h (‘CM’ and ‘CM & PLX’). Simultaneously, but separately, 10 ng ml−1 of IL-1β was added to a portion of the CM collected from the Bowes cells, placed on the HDFs, and incubated for 24 h (‘CM and IL-1β’ and ‘CM and PLX and IL-1β’). RNA was then isolated from each HDF sample and real-time RT–PCR was utilised to analyse (A) CXCR4 and (B) SDF-1 mRNA. *P⩽0.05 and **P⩽0.005.
