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Rather, the depolarized cells started the differentiation process but were restricted from reaching a fully differentiated phenotype, due to instructive signals provided by the changes in Vmem.
The calcium release results were consistent with the ALP production by the cells, as the higher production of the enzyme on the HGL samples indicates that the cells had started the differentiation process but did not proceed with mineralization, producing less calcium overall.
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Fully differentiated cells at day 15 after starting the differentiation process were preincubated with 1 mL nutrition medium lacking fetal calf serum and Dexamethasone for 24 h before starting the experiment.
This is in line with the fact that the knock-down of either of the two transcription factors will decrease the expression levels of the mutual target genes and start the differentiation process [ 4].
This suggests that N-surv is not a marker of KCS in vivo but can identify a population of cells starting the differentiation process, located at the boundary between the basal and suprabasal layers.
The SRY family is especially important in the context of chondrogenesis, as it contains SOX5, 6 and 9, which are essential and sufficient to start the differentiation process [ 23, 34].
Altogether, these results suggest that N-surv marks cells that are starting the differentiation process in normal epidermis, and it is associated with a more aggressive phenotype in human cSCC, possibly contributing to the invasive behaviour of this tumour.
Growth medium was replaced and differentiation medium (including d-Biotin 8 μg/mL, Insulin 0.5 μg/mL, Dexamethasone 400 pg/ml, IBMX 44 μg/mL, L-Thyroxine 9 pg/ml, Ciglitazone 3 μg/mL) was added for 72 h to start the differentiation process.
It is assumed that this cell is selected randomly and that it has started the path of differentiation in the sense that such a cell will never again be selected to reproduce in the stem cell pool.
Therefore, the helper needs to initiate cooperative transmission and starts the priority differentiation phase.
The day before the transfection, cells were plated in six-well plates containing 1% fetal bovine serum in order to start the cell differentiation.
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