Heavy metal toxicity generates reactive oxygen species (ROS) that can contribute to

Overview

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Key numbers

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• prior to imaging, resulting in the less-than-100 % induction rate despite significant oxidative
• supplemented with 10 % fetal bovine serum (ATCC cat. no. 30–2020) and 1 % penicillin/
• a 5 % CO2 atmosphere incubator before the addition of stress inducers. NaAsO2 solution
• (NuPAGE Bis-Tris 4–12 % gel, Invitrogen cat. no. NP0321BOX). Protein was transferred
• a lightbox. Membranes were incubated with blocking buffer (5 % blotting-grade blocker
• 4 % PFA in DPBS at 37 °C for 15 min, and washed three times with DPBS at RT.
• was detected using optics as described in Table S1. Super-resolution images were acquired
• MHz, best dynamic range, and binned to pixels 124–132 in the y-dimension for confocal
• number of cells were imaged for each condition: (DPBS: n = 4 (1 h), n = 3 (24 h); NaAsO2:
• (m1), which is the weighted mean frequency, of the specified spectral region.
• represent 95 % CI (n = 5 biological replicates with 3 technical replicates for each sample).
• G3BP1-positive puncta (n = 3 biological replicates, total of 300 cells were counted). (E)
• sample images used for counting are shown in Fig. S3. Error bars represent 95 % CI (n = 3

Methods (brief)

• cells, acute sodium arsenite (NaAsO2) treatment has long been established as the standard
• 2.1. Biochemical assays on NaAsO2 and H2O2 treatments
• we used IF to determine the specific concentrations of NaAsO2 for 24 h and H2O2 for both 1
• S1), we chose the following four treatment conditions: (NaAsO2) = 500 and 35 μM; (H2O2)
• to these selected treatments. Both chronic treatments of NaAsO2 and H2O2 resulted in the
• 2.2. NaAsO2 and H2O2 induce distinct SG-puncta phenotypes
• Although chronic NaAsO2 and both H2O2 treatments were less effective at inducing puncta
• resulting individual SG-positive cells had more SGs compared to that of either NaAsO2
• 3B). Here, IF images are first collected using an Evident FV4000 confocal laser scanning
• imaging on the same microscope where Raman images are collected (Fig. 4). A fluorescence
• that, in both acute and chronic treatments of NaAsO2 and H2O2, only a subpopulation of
• NaAsO2 treatment, acute treatment with H2O2 and chronic treatment with either NaAsO2
• both durations of H2O2 treatments produced cells with more puncta than either NaAsO2
• collected by centrifugation (500 rcf for 5 min). Pellets were then resuspended in fresh
• a 5 % CO2 atmosphere incubator before the addition of stress inducers. NaAsO2 solution
• were added to sample media the following morning, and cells were incubated at 37 °C for
• ice for 5 min. Adhered cell fragments were collected by scraping, and whole lysate was
• collected. Lysate was centrifuged at 16,100 rcf for 10 min at 4 °C, and the supernatant was

Implications

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Wiki pages this source may touch

• Fish — marine, predatory (tuna, swordfish, shark, king mackerel)
• Fish — marine, non-predatory (sardines, anchovies, salmon, cod)
• Arsenic

Verification notes

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Update history

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