Supplementary data In vitro MIAMI cell migration
Transwell inserts (Millicell cell culture inserts, 8 µm pore size, Millipore) were used to assess MIAMI cell migration in response to factors secreted by Lab1 and U87MG cells. For these experiments, Lab1 and U87MG cells were grown to 70% to 80% confluence in DMEM-HG containing 10% FBS and 1% antibiotics. The media were changed to DMEM with 0.1% BSA and 0.2% antibiotics and cells were cultured for 24 hr. The resulting TCM were centrifuged and stored at -20°C for subsequent use. MIAMI cells (1 x 104) were added to the upper chamber, and 500 µl of control medium (DMEM with 0.1% BSA and 0.2% antibiotics) and Lab1 and U87MG TCM were added to the bottom chamber. After 4 hr, migrated cells on the bottom side of the insert membrane were stained and counted. Data are expressed as percentage of migrated cells.
Supplemental Fig. 1: MIAMI cell migration in response to control medium and Lab1 and U87MG conditioned media. While Lab1 TCM induced no measurable chemotactic response, U87MG TCM led to MIAMI cell migration. (*P < 0.05 versus control medium).
