Methylmercury Uptake into BeWo Cells Depends on

Overview

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Key numbers

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• qPCR (LAT2) from one representative experiment. All three commercially available LAT2 antibodies(Table 1) were proven
• was selected as a multiple of 0.03 µM MeHg, which is equivalent to about 6 µg/L, representing 6 µg/L, representing
• decreased mercury content of the BeWo cells (76% and 58%, respectively) in relation to thecontrols
• mercury content of the BeWo cells (76% and 58%, respectively) in relation to the controls
• transportwaswas5.25.2±±1.7%
• totobasal (3.4±± 1.3%,
• basal(3.4 1.3%, nn == 8). The ratio
• mean values ± SD from two independent experiments. (C) The apical addition of 0.0, 1.5 µM, 1.75 µM,
• from ANOVA are given when p < 0.1. are mean values ± SD from three independent experiments; results
• LAT2 and 4F2hc downregulation resulted in the significantly reduced uptake of leucine (46%
• LAT2 and 4F2hc downregulation resulted in the significantly reduced uptake of leucine (46%
• and 71%, respectively) and methionine (61% and 74%, respectively) when amino acids were added to
• and 71%, respectively) and methionine (61% and 74%, respectively) when amino acids were added
• uptake was observed. The permeability determined by paracellular mannitol transport was 2.1 ± 0.5%
• (n = 6) in experiments examining apical to basal leucine transport. The basal to apical permeability was
• 2.1 ± 0.5% (n = 6) in experiments examining apical to basal leucine transport. The basal to apical
• 4.5 ± 0.9%. With regard to methionine transport, apical to basal permeability was 2.2 ± 0.4% (n = 6),
• permeability was 4.5 ± 0.9%. With regard to methionine transport, apical to basal permeability was
• and basal to apical permeability was 5.0 ± 1.3% (n = 6). The ratio of permeability thus, as expected,
• 2.2 ± 0.4% (n = 6), and basal to apical permeability was 5.0 ± 1.3% (n = 6). The ratio of permeability
• predominantly by the intestinal microflora at a rate of about 1% of the body burden per day and to
• level as commercial LAT2 antibodies (Table 1) were shown to be unable to detect the target protein (20).
• BeWo cells accumulate significantly less mercury (76% and 58%) upon LAT2 and 4F2hc silencing
• of mercury uptake upon LAT2 silencing (75%) but no such effect upon 4F2hc silencing. Moreover,
• uptake of methionine (61% and 74%) and leucine (46% and 71%), whereas LAT1 silencing had no
• on leucine uptake in human primary trophoblast cells (uptake reduced to 87% relative to controls)
• BeWo cells were cultured until 50% confluency on 60 mm dishes (Corning). At this point, the
• 150 mM NaCl, 1% Triton, 0.1% SDS, 0.5% sodium deoxycolate), supplemented with 2 mg/mL aprotinin,
• Blots were blocked for 1 h in 5% nonfat dry milk in tris-buffered saline containing 0.1% Tween
• 20 (TBST), followed by incubation in 5% bovine serum albumin (BSA)/TBST containing the primary
• visualize the signals. For a list of the employed primary and secondary antibodies, see Table 1.
• Table 1. List of the primary and secondary antibodies used in immunoblotting.
• The samples and reference material were acid-digested with nitric acid (69%; Suprapur®;
• by measuring blank test solutions (limit of detection was 0.024 µg/L) and reference materials (Seronorm
• (30.6 ± 7.2 µg/L; n = 19) lay well within the certified range (23.8–55.8 µg/L). All samples were
• measured in duplicate by the working curve method (RSD < 15%).

Methods (brief)

• followedby byaasteady
• and the radioactivity of each cell lysate sample was determined by scintillation counting (TRI-Carb
• The protein samples were separated using SDS-PAGE and transferred to nitrocellulose membranes.
• The samples and reference material were acid-digested with nitric acid (69%; Suprapur®;
• NC, USA). The samples, stabilized with HCl, were stored at 4 ◦ C for up to three days and diluted in
• (30.6 ± 7.2 µg/L; n = 19) lay well within the certified range (23.8–55.8 µg/L). All samples were
• 36. Dalton, P.; Christian, H.C.; Redman, C.W.G.; Sargent, I.L.; Boyd, C.A.R. Differential effect of cross-linking the

Implications

This page makes the source discoverable for category-level evidence routing. Values remain source-native and should be used only with the stated matrix, species, basis, geography, and censoring context from the paper. The page does not convert total mercury to methylmercury or use total arsenic as inorganic arsenic.

Wiki pages this source may touch

• Fish — marine, non-predatory (sardines, anchovies, salmon, cod)
• Shellfish (shrimp, crab, lobster, clams, oysters, mussels)
• Mercury
• Mercury
• Cadmium

Verification notes

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Update history

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