Fig. S5.

Clozapine is not a substrate for hTrxR-1 and hTrx-1.

The effect of 100 μM CLZ on hTrxR-1 and hTrx-1 activities was determined in the presence or absence of 300 μM H₂O₂. The consumption of NADPH was monitored measuring the abs at 340 nm overtime in presence of (A) 10 nM hTrxR-1 and 0.3 mM NADPH and plus (B) 1 μM hTrx-1, as described elsewhere [R. Gencheva, Q. Cheng, E.S.J. Arner, Efficient selenocysteine-dependent reduction of toxoflavin by mammalian thioredoxin reductase, Biochimica et biophysica acta. General subjects 1862(11) (2018) 2511–2517]. In the experiments with H₂O₂, CLZ was preincubated for 10 min at RT with H₂O₂. The concentration of NADPH in the assay mixture was the same in all samples. The higher values recorded at 340 nm in CLZ-treated samples are due to the abs of CLZ, which however remained constant over all the time period of the analysis (not shown). All experiments were performed in quadruplicate, DMSO was used as control. Results represent the mean ± SEM.