1：Experimental Design and Method Selection:

Used single-molecule imaging in living human cells to monitor mRNA localization to SGs and PBs during sodium arsenite-induced stress. Engineered HeLa cell lines with fluorescent markers for SGs (G3BP1-GFP), PBs (TagRFP-T-DDX6), and mRNAs (NLS-MCP-Halo bound to MS2 stem loops).

2：Sample Selection and Data Sources:

HeLa cells were used as the model system. Three mRNA reporters were designed: Renilla luciferase, 5' TOP Renilla (with RPL32 5' UTR), and Gaussia luciferase (ER-associated). Cells were treated with 100 μM sodium arsenite to induce stress.

3：List of Experimental Equipment and Materials:

Microscopes (Zeiss Axioimager Z1, Nikon Eclipse Ti-E with HILO setup), fluorescent proteins (GFP, TagRFP-T, Halo ligands JF549/JF646), antibodies (anti-G3BP1, anti-DDX6, anti-LARP1), siRNAs for knockdown, and various chemicals (sodium arsenite, doxycycline).

4：Experimental Procedures and Operational Workflow:

Cells were induced with doxycycline, treated with sodium arsenite, and imaged over time. Single-molecule tracking, smFISH, immunofluorescence, and western blotting were performed. Data analyzed using Fiji, TrackMate, KNIME, and custom MATLAB scripts.

5：Data Analysis Methods:

Quantified mRNA colocalization with granules, movement dynamics, translation rates (using SunTag system), and degradation rates (using TREAT assay). Statistical analysis with Student's t-test.