Skip to main content
. 2021 Jun 7;10(6):924. doi: 10.3390/antiox10060924

Figure 2.

Figure 2

Reduction of sesamin-induced ARE-GFP expression by depletion of mRNA encoding a Nrf2/Cnc transcription factor or by ectopic overexpression of Keap1. (A,D) GFP fluorescence in brains from adults harboring ARE-GFP reporter (ARE). Brains were prepared from male flies (A1–A4,D1–D4) or female flies (A5–A8,D5–D8). (A1,A2,A5,A6) Brains from control flies with ARE-GFP (ARE-GFP; Act>+) fed on fly diet (A1,A5) or the diet supplemented with 2 mg/mL sesamin (A2,A6). (A3,A4,A7,A8) Brains from flies harboring depletion of cnc in all brain cells using Gal4 driver, Act-Gal4 and ARE-GFP (ARE-GFP; Act > cncRNAi) fed on the fly diet (A3,A7), or the diet with 2 mg/mL sesamin (A4,A8). (D1,D2,D4,D5) Brains from control flies with ARE-GFP (ARE-GFP; elav >+) fed on fly diet (D1,D5) or the diet supplemented with 2 mg/mL sesamin (D2,D6). (D3,D4,D7,D8) Brains from flies harboring ectopic overexpression of Keap1 encoding an inhibitor of Nrf2/Cnc using a pan-neuronal Gal4 driver, elav-Gal4 and ARE-GFP (ARE-GFP; elav > Keap1EY) fed on the diet (Dc3,D7), or the diet with 2 mg/mL sesamin (D4,D8). (B,E) Quantification of GFP intensity of brains from adults harboring ARE-GFP, with or without ubiquitous depletion of cnc (ARE-GFP; Act>+, ARE-GFP; Act > cncRNAi) (B), or brains harboring ARE-GFP with or without pan-neuronal expression of Keap1 from adults (ARE-GFP; elav>+, ARE-GFP; elav > Keap1EY) (E), fed with or without 2 mg/mL sesamin. The intensity of the ARE-GFP fluorescence in adult brains (n ≥ 11) was calculated by Image J for each condition. (C,F) Quantification of GFP mRNA of brains from adults harboring ARE-GFP with or without ubiquitous depletion of cnc (ARE-GFP; Act>+, ARE-GFP; Act > cncRNAi) (C), or brains harboring ARE-GFP with or without pan-neuronal expression of Keap1 (ARE-GFP; elav>+, ARE-GFP; elav > Keap1EY) (F), fed with or without 2 mg/mL sesamin. The relative GFP mRNA level was quantitated by qRT-PCR using total RNA from 40–50 heads for each condition. Scale bars represent 100 μm in the adult brain. ns not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, Student’s t-test.