Fig 2: FasL mRNA decays spontaneously.NC cells and LC cells were exposed to cisplatin in the culture for 48 h to increase the expression of FasL, washed with fresh medium, and then cultured in the presence or absence of LPS (100 ng/ml). The cells were harvested at indicated timepoints and analyzed by RT-qPCR. The curves show the levels of FasL mRNA in LC cells (A) and NC cells (B). C, results of TLR4 RNAi. TLR4d: TLR4-deficient NC cells. #, TLRd NC cells exposed to LPS in the culture. Data are presented as mean ± SEM and represent 6 independent experiments.

Fig 3: Mal deficiency is associated with FasL mRNA decay.A-B, NC cells were prepared and exposed to cisplatin in the culture for 48 h. The cells were then washed with fresh medium and treated with Mal RNAi to knock down Mal expression (A). The cells were cultured in the presence or absence of LPS (100 ng/ml) for 48 h. The bars indicate the levels of FasL mRNA (B). C-D, LC cells were prepared; the cells were transfected with Mal-expressing plasmids or control plasmids to restore the expression of Mal (C). The cells were exposed to cisplatin in the culture for 48 h to up regulate the expression of FasL. After washing with fresh medium, the cells were cultured in the presence or absence of LPS (100 ng/ml) for 48 h. The bars indicate the levels of FasL mRNA in the cells. Data of bars are presented as mean ± SEM. Each dot in bars presents data obtained from one sample. Statistics: t test.

Fig 4: Apoptotic defects in LC cells.The lung cancer (LC) tissues were collected from LC patients (n = 18). LC cells and NC cells (marginal normal tissues; proved by a pathologist) were isolated from the tissues and exposed to cisplatin or saline (vehicle, used as a control) in the culture for 48 h to induce apoptosis. The cells were analyzed by flow cytometry, RT-qPCR and Western blotting. A, gated cells are apoptotic cells. B, summarized data of apoptotic cells in panel A. C, FasL mRNA levels. D, FasL protein levels. E-F, positive correlation between FasL mRNA and apoptotic cells after exposing to cisplatin. Data of bars are presented as mean ± SEM. Each dot in bars presents data obtained from one sample. Statistics: t test.