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Fig. 3. Redistribution of HP1 in immortalized B- and T-lymphocytes. (A) Monocultures, co-cultures and IL-2-treated cells doubly-stained with anti-HP1
(green) and anti-HP1
(red) antibodies. Coarse HP1 foci that appear upon co-culture or IL-2 treatment are more obvious in the HUT-78 line. Notice the convoluted nuclei in HUT-78 monocultures, a typical feature of Sezary cells. Bar, 18 µm. (B) Morphometric data showing the proportion of HP1 and HP1 foci in the nucleus of HUT-78 and RPMI-8226 cells under different culture conditions (blue bars: 0-2 µm foci; green bars: 2-4 µm foci and orange bars: 4-6 µm foci). Optical sections and `projections' of 15 different cells were analyzed in each case. (C) Fold-difference in the abundance of the various classes of HP1 particles upon co-culture or IL-2 treatment in relation to controls (monocultures). The results, shown in a tabular form, were derived from the data presented in B. (D) Western blots showing the levels of HP1, HP1 and histone H3 in nuclear extracts of HUT-78 and RPMI-8226 cells before and after co-culture, or treatment with IL-2.
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