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Twenty-four hours after the last treatments, media is removed and wells are washed with 100 µl of pre-warmed PBS.
After 3 hours transfection media is removed, cells are washed, and fresh growth media is added.
Cells are chilled, media is removed, and the cell layer is rinsed three times with cold PBS.
Only dead/dying cells take up propidium iodide without chemical permeabilization Note that these numbers are probably an underestimation since dead cells tend to float, and some are lost when the culture media is removed.
As with the fractionation assay, when 10% serum media is removed and replaced with 1% serum media and treated with PMA, ERK1/2 is phosphorylated and therefore, activated.
After the final wash, all media is removed from the cells, and transfection cocktails are brought up to a 6 ml volume and added to the culture dishes.
Similar(53)
Neutrophil media was removed and cells washed with fresh RPMI.
Culture media were removed from wells.
After 24 h, culture media was removed from wells.
Afterwards, media was removed and washed twice with PBS.
The media was removed and replaced with fresh media containing GCN and FGCN and incubated for 2 h.
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CEO of Professional Science Editing for Scientists @ prosciediting.com