Figure 6

Metabolic deprivation induced IL-10R. IL-10 decreased PDL1, PD1, FASL, FAS and tumour cell apoptosis on lung ADC cells. (A) Flow cytometric analysis showed that serum starvation or metabolic deprivation (0.4% FCS) induced IL-10R on A549 cells compared with 10% FCS treatment (n=3 per condition). (B) Treatment of serum starved A549 cells with IL-10 leads to an upregulation of pSTAT3 measured by flow cytometric analysis and verified on RNA level by qPCR (n=3 per condition). (C) A549 cells treated with IL-10 showed a significant reduction of apoptosis as shown by decreased Annexin V, a marker of cell apoptosis, under metabolic deprivation compared to the non-treated control cell cultures; (n=3 per condition). (D) FASL, the ligand of FAS (E), a cell apoptosis ligand–receptor, were reduced in A549 by IL-10 treatment under normal metabolic conditions (10% FCS), (n=4 per condition). A549 cells cultured with 10% FCS were treated with IL-10 downregulated the expression of PD1 (F), PDL1 (G) compared with the non-treated control, by flow cytometry (n=4 per condition). Data are shown as mean values±s.e.m. using Student’s t-test *P<0.05, **P<0.01 and ***P<0.001.