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Supplementary LAMP tests using fresh, unpreserved tissues collected during standard parasite passage have demonstrated equal efficacy (data not shown).
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Receptor degradation was indeed prevented by chloroquine (200 μM), as shown by the accumulation of eGFP NPRA in large perinuclear compartments that co-localized with LAMP-1 (Supplementary Figure S1A).
Both provided a 22 °C day/18 °C night, 16 h photoperiod with supplementary light provided by sodium lamps.
Plants were grown under controlled conditions, 13-h photoperiod at a light intensity of 600 mmol.m−2.s−1, with 3-h of supplementary light from metal-halide lamps and using a day/night temperature of 25°C/17°C and 60% relative humidity.
Similar analysis using DNA prepared from whole cells by boiling method indicates that a minimum of 1 cell/reaction was sufficient to give a positive result with the stn LAMP assay under these conditions (Supplementary Figure S5(B)).
Supplementary lighting was provided using sodium vapour lamps.
To ascertain whether spiders in general possess fluorophores, we visually examined abdominal haemolymph in 13 spiders from 10 divergent families (table S1 in the electronic supplementary material) under a 302 nm ultraviolet (UV) lamp.
On the other hand, neither basal lysosomal β-galactosidase (β-gal) activity nor the baseline expression levels of the lysosomal markers LC3-II and lysosomal-associated membrane protein 1 (LAMP-1) correlated well with cellular phenothiazine sensitivity (Supplementary Figures S2A C).
The vesicles stained positive for the endoplasmic reticulum (ER) marker KDEL, but did not stain with the lysosomal marker Lamp-2 or the Golgi marker GM-130 (supplementary material Fig. S1C-E), suggesting glycogen accumulation in the secretory pathway.
There were no changes to the levels of lamp-2a or hsc70 mRNA relative to actin mRNA (Supplementary Figure 3a).
The results presented in Supplementary Figure S2(A) show the co-localization of eGFP NPRA with LAMP-1 after treatment with the lysosomotropic agent ammonium chloride (10 mM).
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