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. 2024 Mar 21;15:2531. doi: 10.1038/s41467-024-46857-w

Fig. 5. The regulatory interplay of NRF2 and GAS41 in transcriptional regulation.

Fig. 5

a RT-qPCR analysis of TIP60, TRRAP, SLC7A11, NQO1, and GCLC mRNA levels in A549 cells transfected with control siRNA (siCTRL), TIP60 siRNA (siTIP60), or TRRAP siRNA (siTRRAP). b ChIP-qPCR analysis of CBP binding on SLC7A11 and GCLC promoter region in A549 sgGAS41 cells transfected with NRF2 along with vector or GAS41 WT. c ChIP-qPCR analysis of overexpressed NRF2 binding on SLC7A11 and GCLC promoter region in A549 sgGAS41 cells transfected with NRF2 expressing plasmid along with vector, GAS41 WT, GAS41 W93A mutant, or GAS41 L211A mutant. d ChIP-qPCR analysis of overexpressed GAS41 enrichment on GCLC and SLC7A11 promoter region in A549 shNRF2 Tet-On inducible cells pre-incubated without or with doxycycline (0.2 μg/mL) for 48 h, then transfected with HA-tagged GAS41 expressing plasmid along with vector or Flag-tagged NRF2 expressing plasmid. e, f Snapshot of NRF2 CUT&RUN signal in A549 sgNC and sgGAS41 cells at GCLC (e) and NQO1 (f) genes loci. g ChIP-qPCR analysis of NRF2 binding on SLC7A11 and GCLC promoter region in A549 sgNC and sgGAS41 cells. h ChIP-qPCR analysis of GAS41 enrichment on SLC7A11 and GCLC promoter region in shNRF2 A549 without or with doxycycline (0.2 μg/mL) for 72 h. i Western blot of GAS41, NRF2, SLC7A11, and GCLC protein levels in H1299 treated with DMSO or tBHQ (50 μM) for 24 h. j ChIP-qPCR analysis of GAS41 enrichment at SLC7A11 and GCLC promoter region in H1299 treated with DMSO or tBHQ (50 μM) for 24 h. k Work model for the role of GAS41 in antioxidant transcription regulation through anchoring NRF2 with histone acetylation. Created with BioRender.com. Data are mean ± SD of n = 3 independent biological repeats. p values were calculated using unpaired, two-tailed Student’s t test. The experiment (i) was repeated three with similar results and representative results are shown. Source data are provided as a Source Data file.