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Ulose paperDNA hybrid structures have been formed by soaking 1 5-mm wide precision
Stretching was performed at a price of ca. 3 mm min-1. Temporal Laquinimod Autophagy recovery experiments. For the temporal recovery experiments, the DNA hydrogels had been constantly imaged for about 100 s right after gel rupture. The ratios of red to green fluorescence had been measured on the same compact location as for the stretching experiment and have been normalized to the minimal (just before stretching) and maximal (straight away before rupture) fluorescence ratio. Image treatment and evaluation. The series of fluorescence photos were treated applying ImageJ by initially performing a background cleaning (200 pixel radius). Then, the temporal profiles of each the red and green channels in the area of interest had been extracted. The values plotted correspond towards the redgreen ratio after subtraction of your initial RG ratio. The strains were calculated based around the sample lengths measured manually every five frames of the videos. We systematically measured the fluorescence ratios on a small location in the center from the tensile specimen (in yellow), which focuses on a section in the sample that is definitely really stretched and grants maximal sensitivity (Supplementary Fig. 6a ). We also measured the fluorescence more than the full location (in purple), which confirms the overall improve of red fluorescence and the absence of bleaching because the amount of green fluorescence remains constant during the whole stretching experiment (Supplementary Fig. 6b). This technique enables acquiring reproducible and quantitative strainfluorescence curves for all systems as presented in Supplementary Fig. 6d for the diverse modules, at the same time as for the temporal recovery experiments (Supplementary Fig. 6h).Data availabilityThe data that assistance the plots inside this paper along with other findings of this study are accessible in the corresponding author upon reasonable request.Received: 12 July 2018 Accepted: 9 January
ARTICLEhttps:doi.org10.1038s41467-019-10648-OPENThe Polycomb protein Ezl1 mediates H3K9 and H3K27 methylation to repress transposable components in ParameciumAndrea Frapporti 1,6,7, Caridad MirPina 1,7, Olivier Arnaiz 2, Daniel Holoch three, Takayuki Kawaguchi Adeline Humbert1, Evangelia Eleftheriou2, B ang e Lombard4, Damarys Loew four, Linda Sperling two, Karine Guitot 5, Rapha Margueron3 Sandra Duharcourt1234567890():,;1,In animal.Ulose paperDNA hybrid structures have been formed by soaking 1 5-mm wide precision Whip dust-free paper (Kimberly ClarkTM; Kimtech Science) and submitting it to the same thermal therapy beneath a polyurethane cover to prevent water evaporation. For the composite, 5 m sulfate-functionalized polystyrene microspheres (10 wt in water) had been mixed in 1: 1 ratio with all the liquid hydrogel premix at 1.six wt and placed directly within the sampleholder, followed by a temperature ramp for setting the hydrogel before mechanical tests. Tensile tests with in situ fluorescence monitoring. Imaging was performed working with a AxioZoom widefield microscope using a 1 objective lens (Aperture 0.25) sequentially imaging the green channel (Filter set 38 HE: excitation 440470, emission 525550; exposure 200 ms) and also the red channel (Filter set 63 HE: excitation 572525, emission 629662; 500 ms exposure) resulting in 0.44 pictures with filter alterations. Stretching was performed utilizing a screw-driven custom-built extensometer permitting the DNA hydrogel to become immersed in TE buffer supplemented with 100 mM NaCl and 12 mM MgAc2 through the experiment. The buffer solution was regularly changed with ice-cooled fresh buffer to maintain the sample below 20 .
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