Fig. 6

Gremlin1 depletion may affect 66cl4’s ability to extravasate. (a-c) Analysis of the tumor cells ability to penetrate a confluent monolayer of human umbilical vein-endothelial cells (HUVECs) using the xCELLigence Real-Time Cell Analysis (RTCA) Systems. Representative graphs depicting the change in normalized cell index after addition of tumor cells or conditioned medium. Pure growth medium was used as a control. (a) 67NR and 66cl4. (b) Conditioned medium of 67NR and 66cl4. (c) 66cl4 non-target control and two Grem1 knockout clones. Results are shown as mean ± SD (A-C). (d) Representative image of 6 dpi zebrafish larvae showing the invasion of 66cl4 wildtype cells, non-target control (NT_mix) and Grem1_1.3 (red). The vasculature is shown in green. (e) Quantification of invasive cluster numbers in 66cl4 injected zebrafish larvae. Probability represents the percentage of zebrafish larvae in which tumor cell extravasation was observed. Results are shown as mean ± SEM. The line indicates mean cluster number. Student’s t-test, *0.01 < P < 0.05