1：Experimental Design and Method Selection:

The study used in vitro experiments with 661W cone cells and in vivo experiments with rd10 and rd10/nrf2-/- mice to investigate Sig1R activation effects on NRF2 and oxidative stress. Methods included cell viability assays, immunodetection, qRT-PCR, western blotting, KEAP1-NRF2 binding assays, NRF2-ARE binding assays, siRNA transfection, ERG, SD-OCT, and histological analyses.

2：Sample Selection and Data Sources:

661W cone cells were used as an in vitro model. Mice included rd10 (B6.CXBI-Pde6βrd10/J) and nrf2-/- mice on a C57BL/6 background. Data were sourced from cell cultures and animal models.

3：CXBI-Pde6βrd10/J) and nrf2-/- mice on a C57BL/6 background. Data were sourced from cell cultures and animal models. List of Experimental Equipment and Materials:

3. List of Experimental Equipment and Materials: Equipment included fluorescence microscopes (Axioplan-2, Carl Zeiss), plate readers (Synergy H1 Hybrid Multi-Mode, BioTek Synergy 2), luminometers (Lmax11384, Molecular Devices), ERG apparatus (custom-made with Psychlab EEG8 amplifier, National Instruments NI-6229 digitizer), SD-OCT system (Bioptigen Envisu R2200), and confocal microscopes (Zeiss LSM 780). Materials included DMEM, FBS, antibodies (e.g., anti-Sig1R, anti-NRF2), assay kits (e.g., KEAP1-NRF2 inhibitor screening assay kit, TransAM NRF2 kit), and chemicals like (+)-PTZ and tBHP.

4：0). Materials included DMEM, FBS, antibodies (e.g., anti-Sig1R, anti-NRF2), assay kits (e.g., KEAP1-NRF2 inhibitor screening assay kit, TransAM NRF2 kit), and chemicals like (+)-PTZ and tBHP. Experimental Procedures and Operational Workflow:

4. Experimental Procedures and Operational Workflow: Cells were cultured and treated with (+)-PTZ or tBHP. Viability was assessed using MTT assay. Immunodetection, qRT-PCR, and western blotting were performed to measure protein and gene expression. Binding assays used fluorescence polarization and luciferase reporters. In vivo, mice were treated with (+)-PTZ and subjected to ERG, SD-OCT, and histological analyses post-euthanasia.

5：Data Analysis Methods:

Data were analyzed using GraphPad Prism and Igor Pro software. Statistical tests included t-tests, ANOVA with post-hoc tests (Holm-Bonferroni, Tukey's, Newman-Keul's). Fluorescence intensity was quantified with NIH ImageJ.