Figure 2. Palmitoylated cld7 supports anchorage independent growth and metastasis formation.

(A) Wt, kd and rescue ASML cells (500) were seeded in soft agar in 10 cm Petri-dishes. Colonies were counted after 14 d. The mean number ± SD (triplicates) of colonies and representative examples are shown. (B, C) BDX rats received 1 × 10⁶ wt, kd or rescue ASML cells, ifp. (B) The number of surviving rats and the mean survival time are shown; (C) the number of visible lung metastases, the mean draining lymph node diameter, the number of tumor cell colonies growing in suspended lung and LN tissue was evaluated after 4 wk. (D) Shock frozen lung sections from ASML wt, -cld7^kd, cld7^mPalm and -EpC^resc bearing rats were stained with anti-EpC, -cld7 and -CD44v6 (scale bar: 120 μm). Tumor cells (EpC⁺, cld7⁺, CD44v6⁺) are only seen in the lung of ASML^wt and -EpC^resc bearing rats. In the lungs of ASML-cld7^kd and -cld7^mPalm bearing rats only bronchiolar epithelium (*) is stained by anti-EpC and anti-cld7. Anchorage-independent growth and metastasis are severely impaired in both kd clones, but are regained in ASML-EpC^resc and partly ASML-EpC^mAG, but not in -cld7^mPalm cells, indicating only palmitoylated cld7 supporting metastasis.