Fig. 3: Treatment with 20 µM 1,4 dihydroxy quininib affects ROS, biliverdin, GSH, LOOH levels and NRF2, GPX4, GCLM expression in OMM2.5 cells. | Cell Death Discovery

Fig. 3: Treatment with 20 µM 1,4 dihydroxy quininib affects ROS, biliverdin, GSH, LOOH levels and NRF2, GPX4, GCLM expression in OMM2.5 cells.

From: 1,4-dihydroxy quininib activates ferroptosis pathways in metastatic uveal melanoma and reveals a novel prognostic biomarker signature

Fig. 3

A Evaluation of the effects of 20 µM Q7 on ROS levels in OMM2.5 cells after 8 (n = 4 independent experiments) and 24 (n = 3 independent experiments) h of treatment (*p < 0.05). B Western blot analysis of NRF2 expression in 0.5% DMSO or 20 µM Q7 treated OMM2.5 cells after 4 (n = 6 independent experiments), 8 (n = 6 independent experiments) and 24 (n = 5 independent experiments) h of treatment (*, p < 0.05; ** p < 0.01). C Densitometric quantification of NRF2 vs beta-actin as determined by at least three independent western blot experiments as in B. The results are expressed as means ± SEM. D Measurement of intracellular biliverdin in 0.5% DMSO- or 20 µM Q7- treated OMM2.5 cells after 4, 8 and 24 h of treatment (** p < 0.01). N = 3 independent experiments. E Western blot showing GPX4 expression in 0.5% DMSO- or 20 µM Q7- treated OMM2.5 cells after 4 (n = 3 independent experiments), 8 (n = 4 independent experiments) and 24 (n = 7 independent experiments) h. F Densitometric quantification of GPX4 vs beta-actin as determined by at least three independent western blot experiments as in E (*p < 0.05; ***p < 0.001). G Measurement of non-protein thiol group (RSH) concentrations after 4 (n = 4 independent experiments), 8 (n = 4 independent experiments) and 24 (n = 3 independent experiments) h of treatment (**p < 0.01; ***p < 0.001). H Evaluation of LOOH levels after 4 (n = 4 independent experiments), 8 (n = 4 independent experiments) and 24 (n = 3 independent experiments) h of treatment (**, p < 0.01). The results are expressed as means ± SEM. I Western blot showing GCLM expression in 0.5% DMSO- or 20 µM Q7- treated OMM2.5 cells after 4 (n = 6 independent experiments), 8 (n = 5 independent experiments), and 24 (n = 4 independent experiments) hours. J Densitometric quantification of GCLM normalized to beta-actin as determined by at least three independent western blot experiments as in I (** p < 0.01). The histograms report mean ± SEM. Q7 = 1,4-dihydroxy quininib; h = hours.

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