Figure 5. Cld7 and apoptosis resistance.

(A) Wt, kd and rescue ASML cells were cultured in the presence of increasing amounts of cisplatin. Flow cytometry analysis of the percent of AnnV+, AnnV+PI+ and PI+ cells; mean ± SD (triplicates) are shown, significant differences to wt cells: *. (B) WB of Akt, Pten, pPten and Pten phosphorylating kinases in untreated and PMA-treated wt, kd and rescue ASML cells; (C) sucrose gradient fractions of the lysates as above and WB with anti-Pten, -pPten, -GSK3β, -CK2, -src, -cld7 and -Tspan8; (D) Membrane and cytosol lysates from cells as above were blotted with anti-GSK3β, -CK2, -src, -pPten and -cld7. (E) WB with anti-Pten, -cld7, -EpC, -GSK3β, -CK2 and -src after precipitation with anti-GSK3β, anti-CK2 and anti-src. Drug resistance of ASML-cld7^kd cells is severely impaired and not rescued in ASML-cld7^mPalm cells. Upregulation of Pten in PMA-treated ASML-cld7^kd and -cld7^mPalm cells indicates Pten repression only by palmitoylation-competent cld7. Concomitant pronounced cytoplasmic Pten phosphorylation does not contribute rescuing apoptosis resistance.