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Ana-1 cells were seeded in 6-well plates at a density of 2 × 106 cells per well in quadruplicate and grown to confluence for 24 h.
Interestingly, even in the complete absence of differentiation medium, Tsc2−/− MEFs incubated at confluence for 7 days in normal growth medium accumulated more lipid than differentiated Tsc2+/+ cells (Figure 2, panels A and B).
Cells were grown to confluence for 48 h in six-well plates.
The conditioned media were obtained after incubation of C9 cells at confluence for 48 hours in the presence of either chondrogenic medium or osteogenic medium.
For cell cycle entry assays, HFFs were first driven into quiescence by maintaining cells at confluence for 3 days in medium containing 10% FBS followed by 3 days of culture in DMEM containing 0.1% FBS.
The colonies showing a reduction of GUS expression were cultured in order to reach 70%-80 70%-80nfluence for eight passages.
Small portions from mycelium were removed under stereomicroscope and cultured on MM medium (as described above) in order to reach 70%-80 70%-80nfluence for eight passages.
Flow-cytometric analyses showed that the treatment of CoCl2 and SIN-1 for 24 h in confluence state of hADSCs increased the proportion of subG1 phase cells, and that miR-302d transfection decreased the subG1 population in the presence of CoCl2 and SIN-1.
For detailed procedures, see the KFS Access Requests page in Confluence.
For additional information visit the KFS Access Requests page in Confluence.
Confluence's mashup of activity streams and task management is a noble experiment, but I expect it will only work out well for companies where most employees already spend a lot of time in Confluence.
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Since I tried Ludwig back in 2017, I have been constantly using it in both editing and translation. Ever since, I suggest it to my translators at ProSciEditing.

Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com