Figure 4. Palmitoylated claudin7 supports invasiveness and protease activity.

(A) Wt, kd and rescue ASML cells were seeded on matrigel. Matrigel invasion and penetration was evaluated after 16 h incubation. The mean percent ± SD of invading and penetrating cells and representative examples are shown. (B) Zymography of culture supernatants of ASML^wt, -cld7^kd and -cld7^mPalm cells; MMP2 and MMP9 bands are indicated. (C) Lysates from cells as in (A) were precipitated with anti-CD147 (EMMPRIN), anti-uPAR and anti-MMP14 and were blotted with anti-cld7 or were precipitated with anti-cld7 or anti-EpC and blotted with anti-uPAR and anti-CD147. WB of lysates with anti-uPAR, -MMP14 and -CD147 are included as controls. (D) Cells as above were stained with anti-CD147 (green) and either anti-EpC (red) or anti-cld7 (red). Digital overlays are shown (scale bar: 10 μm). ASML-cld7^kd cells are poorly invasive and invasiveness is not restored in ASML-cld7^mPalm cells. This also accounts for reduced MMP9 activity and fits to reduced coimmunoprecipitation of cld7^mPalm with MMP14 and uPAR. EpC has a weaker impact on invasiveness and MMP activity.