Fig. 22

FL496, a FL118 analogue, exhibited extended target specificity: A Effects of FL118 and FL496 on the expression of NRF2 in UOK262 tumor cells. Sub-confluent cells grown in complete medium in 6-well plates were treated with FL118 and FL496 for 16 h at 0, 10, 100 and 500 nM, respectively as shown. Cells were then analyzed with Western blots using the NRF2 antibody. GAPDH is the internal protein loading control. B Effects of FL496 on the modulation of potential molecular targets in the Type 2 pRCC cell line, FHpRCC UOK262. Sub-confluent cells grown in complete medium in 6-well plates were treated for 16 h with FL496 at the concentration of 0 nM, 10 nM, 100 nM and 500 nM as shown. Cells were then lysed for Western blot analyses with antibodies for the corresponding molecular protein targets. GAPDH is the internal protein loading control. C FL496 is not an ABCG2 efflux pump substrate: HEK293 cells stably transfected with either pcDNA3 empty vectors (control) or with pcDNA3-ABCG2 expression vectors were treated with or without a series of concentrations of FL496 for 72 h as shown. Cell viability was then analyzed using MTT assay. Each bar is the mean cell growth/viability ± SD derived from 5 independent parallel testing. D FL496 mechanism of action model based on the data observed (A, B), which was adapted from the publication entitled “Targeting ABL1-Mediated Oxidative Stress Adaptation” [192].