Fig 4. AS1842856 leads to a transition from quiescence to the G₁ phase of the cell cycle of T lymphocytes.

(A) PBT were cultured for 7 days in the presence of AS1842856 (500nM) or vehicle only. RNA and DNA cell content was measured by FACS on the gated cells after acridine orange staining. Untreated T cells activated for 3 days with anti-CD3/CD28 beads were used as a positive control of cells increasing both their RNA and DNA content. Result obtained with one representative donor (left panel) and mean results +/- SE with cells from 7 independent donors (right panel) are shown. (B) PBT from 3 independent donors were cultured as in A, then collected, lysed and immunoblotted using specific antibodies against p27, Cdk2 and SamHD1 proteins. The same lysates were also analyzed for Rb and SAMHD1 phosphorylation with phospho-specific antibodies.