Supplemental Figure
-
Fig. S1. Cell
cycle arrest in human primary fibroblast in response to iron depletion. (A) In
vitro replication reactions of nuclei isolated from HFF cells treated with the
iron chelator mimosine for 24 hours (mim) or from HFF cells that underwent the
same treatment followed by release from the arrest by the removal of mimosine
for 4 hours (mim 4 h rel). Nuclei were incubated in an elongation buffer
(åbuffer?, light grey bars) or in the same buffer supplemented with 100 mg HeLa cytosolic extract (åcytosol?, dark
grey bars). Percentages of nuclei incorporating labelled nucleotides were
determined. (B) HFF cells were kept in mimosine for 24 hours (0 hour sample),
and then either mimosine was removed by replacing the medium (mimosine release,
rel), or 1 mM FeSO4 (Fe) was added for the indicated time periods.
The presence of DNA breaks is indicated by a focal pattern of H2AX
phosphorylation (green). Representative g-H2AX
containing nuclei are shown.
