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. 2012 Jun 13;7(6):e38102. doi: 10.1371/journal.pone.0038102

Figure 2. APC depletion increases chromatin compaction in HeLa cells.

Figure 2

Asynchronously growing HeLa cells stably expressing GFP-H2B and mCherry-H2B were depleted of APC using siRNA (siAPC) or transfected with non-targeting siRNA (siContr), and donor fluorescence lifetime was measured using time-correlated single photon counting technique. A . Normalised lifetime values for APC- (grey bars) and mock- (white bars) depleted interphase cells (left) and mitotic cells (right). P-values are given underneath the plots. Please note that APC-depleted mitotic cells display significantly shorter FRET lifetime, indicating a higher degree of chromatin compaction. At least ten cells were measured for each condition. B . Level of APC depletion of cells used in A is visualized by immunobloting the corresponding lysates with anti-APC antibodies. Tubulin is used as a loading control. C . Representative chromatin images from control (left) and APC-depleted (right) mitotic cells. FRET efficiency is shown in false colors, with blue corresponding to low FRET efficiency (i.e. less compaction), and red corresponding to high FRET efficiency (i.e. more compaction).