Download PDF by Reti L.: Beta-Phenethylamines Manske and Holmes, The Alkaloids

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36 Tasdemir et al. lamp-1 lamp-2 Lysosome-associated membrane protein 1 Lysosome-associated membrane protein 2 NM_002294 NM_005561 NM_002647 sense 5’-GGCUGAAACUACCAGUAAAdTdT-3’ antisense 5’-UUUACUGGUAGUUUCAGCCdTdT-3’ sense 5’-GGAGGCAAAUAUCCAGUUAdTdT-3’ antisense 5’-UAACUGGAUAUUUGCCUCCdTdT-3’ sense 5’-CGAGAAAUGCAACACGUUAdTdT-3’ antisense 5’-UAACGUGUUGCAUUUCUCGdTdT-3’ sense 5’-GGAAUCCAGUUGAAUACAAdTdT-3’ antisense 5’-UUGUAUUCAACUGGAUUCCdTdT-3’ sense 5’-GCUGUGCGGUCUUAUGCAUdTdT-3’ antisense 5’-AUGCAUAAGACCGCACAGCdTdT-3’ sense 5’-GCGGUCUUAUGCAUUGGAAdTdT-3’ antisense 5’-UUCCAAUGCAUAAGACCGCdTdT-3’ (12) (12) (82) Notes: siRNAs are provided either as 100 mM stock solutions (to be stored as such at −20°C), or in dehydrated form.

Then use the wet spatula to transfer the gelatin and cells to a Petri dish on ice, and cut the cell pellet to small cubes/sticks (maximum side length about 2 mm). 20 Eskelinen Before freezing, the cells need to be incubated in a cryoprotectant. 7 M sucrose–15% PVP) are mostly used. PVP gives plasticity to the block and thus helps cryosectioning. Incubate the cell cubes in sucrose-PVP at 4°C, with occasional mixing, for several hours to overnight, until the cubes sink to the bottom. Trim the cubes under a stereo microscope and mount on cryoultramicrotome holders at 4°C (in a cold room, or using an ice bath).

Cell Biol. 25, 1025–1040. 4. Lum, J. , Bauer, D. , et al. (2005) Growth factor regulation of autophagy and cell survival in the absence of apoptosis. Cell 120, 237–248. 5. , et al. (2004) The role of autophagy during the early neonatal starvation period. Nature 432, 1032–1036. 6. Eskelinen, E. , and Saftig, P. (2003) At the acidic edge: emerging functions for lysosomal membrane proteins. Trends Cell Biol. 13, 137–145. 7. , et al. (2000) Accumulation of autophagic vacuoles and cardiomyopathy in LAMP-2 -deficient mice.

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Beta-Phenethylamines Manske and Holmes, The Alkaloids by Reti L.


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