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Fig. 3. Lamin B1 depletion correlates with altered morphology in nucleoli and nuclear speckles. Cells co-transfected with RNAi vectors and a vector expressing histone H2B were analysed after 48 hours to monitor the morphology of major nuclear compartments. (A) Sites of pre-rRNA synthesis in nucleoli were visualised by indirect immunolabelling of fibrillarin (green). Changes were monitored in cells expressing DsRed-histone H2B (red) and different patterns recorded; shown in cartoon form and occurrence as a frequency histogram [LB1 kd, 187 cells (four experiments); Control, 100 cells (two experiments)]. Control cells were transfected with empty vector, although untransfected cells in RNAi samples also serve as controls. (B) Nuclear speckles were visualised by indirect immunolabelling of the splicing factor SC-35 (green). Structural changes were monitored in cells expressing DsRed-histone H2B (red) and the appearance and frequency of major changes described (LB1 kd, 196 cells (five experiments); Control, 98 cells (two experiments). (C) The structure of the chromatin compartment in HeLa cells was monitored by expression of DsRed-histone H2B. Typical examples of untreated cells (control) and cells with lamin A/C or B1 depleted (Kd LA/C; Kd LB1) are shown. Scale bars: 5 µm (A,C); 10 µm (B).
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