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(e) Representative phase contrast photomicrographs of endothelial tube formation of HUVECs cultured on Matrigel with 1X or 10X RMS-derived exosomes from the specified cell lines, as well as control (Exo Free medium), as specified.
All additions were made to this medium, as specified in the figure legend.
Exponentially growing human fibroblasts were transfected with 1 µM of morpholino antisense oligonucleotide targeting top2α mRNAs (MO-Top2a; 5'-TTTACAGGCTGCAATGGTGACACTT-3'; Gene Tools) or a standard control (MO-SC; 5'-CCTCTTACCTCAGTTACAATTTATA-3'; Gene Tools) using 6 µM of Endo-Porter reagent (Gene Tools) in complete medium, as specified by manufacturers.
Agonist was added to the same medium as specified in figure legends.
Twelve hours later, mice were challenged with a 10 μL inoculum containing 1 or 0.1 ID50 of HSV-2 in Bartels medium as specified below.
Where indicated, metformin (Sigma; stock solution: 300 mM in water), tunicamycin (Sigma; stock solution: 10 mg/ml in DMSO, final concentration 100 ng/μl) and staurosporine (Sigma; stock solution 1 mM in DMSO) were added to the medium as specified in the Results section.
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The interaction effects (Eta squared η2) were in the medium effect range as specified in Cohen (1977) (η2 = .01, η2 = .06, and η2 = .14 as small, medium, and large effect sizes, respectively).
After 24 hr, medium was changed as specified and subsequently refreshed every other day.
The culture medium may change as specified in each cultural condition, together with other important growth factors, such as initial pH level, culture temperature and so on.
This was followed by two washes with LM on ice, mounting on coverslips in LM medium containing inhibitors as specified and subsequent observation by CLSM.
Based on the above researches, it is necessary to perform detailed atomic simulation to get diffusion coefficient of the particular anticancer drug in the specified medium as accurate as possible.
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