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CDK6 forms a complex with cyclin-D and targets the retinoblastoma protein, allowing passage through the G1/S phase restriction point.
Corroborating these data, results shown in Figure 2d revealed that apigenin led to a complete degradation or inhibition of expression of CDK6, a protein essential for passage through the G1/S phase restriction point.
Together, these observations demonstrated that both primary and tumor cells exclusively generate mono-phosphorylated Rb during all of early G1 phase before being converted in a quantum step to hyper-phosphorylated Rb at the late G1 phase Restriction Point.
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To study whether greater number of cell division is due to higher number of cells entering cell cycle, we used BrdU incorporation assay to assess the number of cells entering the S phase, the restriction point, of cell cycle.
Cyclins D and CDKs are key determinants in the passage of the cell cycle through G1 phase, past the restriction point and into S-phase.
More specifically, this sensitive period includes the mid-to-late G1 phase culminating at the Restriction point (or R point), and the onset of DNA replication.
The cyclin D3, CDK4 and 6 activities in the mid-late G1 phase control the G1 restriction point and activation of the cyclin E/CDK2 complex (Planas-Silva and Weinberg, 1997).
Cyclin D family accumulation is needed for passage through the restriction point in G1 phase, just before entry into S phase 19.
In cycling cells, E2F1 and cyclin E expression normally reaches a maximum in late G1/early S-phase after passing through the Restriction point.
Cell senescence is a biochemical process exhibited by metabolically active cells whose cell cycles are frozen beyond the restriction point in G1 phase.
If the latter were involved, then one would expect a concomitant increase in the expression of the CDK inhibitors p21 and p27, which negatively regulate progression through restriction point in G1 phase.
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