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. 2017 Aug 2;18(8):1680. doi: 10.3390/ijms18081680

Figure 3.

Figure 3

NRF2 knockdown sensitises LTTs towards cisplatin by increasing DNA damage. NRF2 protein (a) and mRNA (b) expression was measured in LTTs after siNRF2-mediated knockdown and their negative controls. As a loading control, α-Tubulin was detected. GSR, NQO1, HMOX1, GPX1, GSTP1, and p62/SQSTM1 mRNA expression in siNRF2-transfected (c) RT-112-LTT, (d) J82-LTT, (e) 253J-LTT, (f) T-24-LTT and their negative controls were measured by qRT-PCR. Expression levels in the negative control were set as 1. SDHA mRNA was used as a reference and relative expression was calculated by the 2−ΔΔCt method. (g) Cell viability was measured 72 h after cisplatin treatment by CellTiterGlo assay in LTTs transfected with siRNA targeting NRF2 or a non-targeting negative control siRNA. (h) Quantification of pH2A.X Ser139 foci by immunofluorescent staining in LTTs transfected with siRNA targeting NRF2 or a non-targeting negative control siRNA and treated or not treated with cisplatin after 72 h. Values represent the mean ± SD of biological duplicates, * p < 0.05.