LAM cell medium is chemotactic for fibroblasts, and this is partially dependent on CXCR4. (a) 621–101 cells secrete CXCL12 protein; this is partially sensitive to rapamycin. (b) The presence of 621–101 cell conditioned medium (CM) increases fibroblast migration, which is partially blocked by AMD3100. (c) Fibroblasts migrate towards CXCL12 in a modified Boyden chamber (Transwell) assay; this is inhibited by the CXCR4 receptor antagonist AMD3100 at 100μg/ml (*p = 0.05). (d) Fibroblasts migrate towards 621–101 cells in a bidirectional migration assay. (i) Removal of the fence leaves a gap between the peripheral fibroblasts (green) and the central reservoir of (red) 621–101 cells (dotted), (ii) Fibroblasts migrate towards the central reservoir (arrowed), (iii) fibroblasts migrate into the central area (arrowed). (e) Addition of 200μg/ml AMD3100 reduces migration of fibroblasts in response to 621–101 cells by 56%. In the absence of 621–101 cells in the central reservoir migration is reduced by 93%.
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