Fig. 6
Mitochondrial ROS scavenging prevents LAP-induced up-regulation of ABC transporters and chemo-immuno-resistance. NCI-H2228 cells were cultured as indicated in Fig. 1a. When indicated, 50 μM elesclomol (Es) or 0.4 μM mitoquinol (mQ) were added to the normoxic or hypoxic cultures. a-b ChIP of C/EBP-β on ABCB1, ABCC1 and ABCA1 promoter (a) and ABCB1, ABCC1, ABCA1 mRNAs (b), measured by RT-PCR, in technical triplicates. Data are means±SD (n = 4 biological replicates). *p < 0.05,**p < 0.01,***p < 0.001: H, H/N, H/N/H versus N cells; °p < 0.05 °°°p < 0.001: Es/mQ-treated cells versus untreated cells. c After normoxic and hypoxic cultures, cells were treated for 48 h in normoxia (20% O2) with increasing concentrations (from 1 × 10− 9 to 1 × 10− 5 M) of cisplatin (Pt). Cell viability was measured in technical quadruplicates (n = 3 biological replicates). Representative (inhibitor) vs. normalized dose-response curves and relative IC50, obtained with the GraphPad Prism 9 software. d-e Percentage of Ki67+IFN-γ+ Vγ9Vδ2 T cells (d) and percentage of annexin V+PI+ NCI-H2228 cells (e), as index of tumor cells killed by Vγ9Vδ2 T-cells, collected after the co-cultures with the NCI-H2228 cells, evaluated by flow cytometry, in technical duplicates. Data are means±SD (n = 5 biological replicates). ***p < 0.001: H, H/N, H/N/H versus N cells; °°°p < 0.001: mQ-treated cells versus untreated cells