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Micro-irradiation by UV-laser (355nm). GFP BMI1-U2OS (green) cells were micro-irradiated by UV laser (355 nm) and the induction of DSBs was verified using antibody against ?H2AX (red strip). Bar represents 4 ?m.
Recruitment of HP1? to UV damaged chromatin is influenced by the histone acetylation state. (A) Live control GFP HP1?-3T3 cells were micro-irradiated and increased accumulation of HP1? at damaged chromatin was detected within ~200 s. TSA prevented HP1? recruitment to damaged chromatin, but HP1? was not absent at micro-irradiated regions of TSA treated cells. Bar represents 4 µm. (B) FRAP was performed in UV-damaged chromatin (micro-irradiated euchromatin and heterochromatin) of control and TSA-treated GFP HP1?-3T3 cells. (a) A comparison HP1? recovery at heterochromatin of control and TSA treated cells. (b) HP1? recovery at euchromatin of control and TSA treated cells. (c) A comparison of HP1? recovery at heterochromatin and euchromatin of control cells. (d) A comparison HP1? recovery at heterochromatin and euchromatin of TSA treated cells.
Levels of Oct4 and Nanog proteins within GOWT1 mESC colonies. (A) GOWT1 cells expressing high levels of Oct4 had low levels of Nanog protein (see magnification in upper panel). Nanog protein was preferentially abundant in the cells that occupied the interior of the colony (lower right panel), but there was no correlation for the cells with high and low Oct4 levels (lower left panel). The Oct4 pattern was studied in living cells, but Nanog distribution was acquired after fixation of the same colony by paraformaldehyde; thus, a slight shift in colonies can occur.