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. 2021 Dec 10;45(3):134–147. doi: 10.14348/molcells.2021.0074

Fig. 6. KKC080096 activates Nrf2 pathway and upregulates the expression of Nrf2 target enzymes in microglial cells.

Fig. 6

(A) BV-2 cells were treated with various concentrations of KKC080096. After 24 h, the cells were harvested and the cell lysates were subjected to western blotting for Nrf2 using β-actin as an internal control (upper panel). After 6 h, the cells were harvested and the nuclear and cytosolic fractions were subjected to western blotting for Nrf2, lamin B (a nuclear marker used as an internal control), and HSP90 (a cytosolic marker used as a negative control) (lower panel). (B) Surface plasmon resonance was recorded in the presence of KKC080096. The data are expressed as resonance units (RU). (C) The biotinylated Nrf2 ETGE peptide immobilized on streptavidin-agarose beads was incubated with Keap1-Kelch domain in the presence of KKC080096, and the Nrf2-bound Keap1 was detected using western blotting. The data are plotted as percentage of control ± SEM; **P < 0.01, ***P < 0.001. (D) BV-2 cells were treated with various concentrations of KKC080096 for 6 h. The cells were harvested and RT-PCR was performed for NQO1, GCLC, and GCLM. (A and D) The number on each gel pictogram represents the fold increase with respect to untreated control, calculated from densitometry and normalized against respective loading control.