Fig. 11

RNAi-mediated depletion of MRG15, Tip60, and YETI in S2 cells affects mitosis and cytokinesis. Examples of mitotic defects found in MRG15, Tip60, and YETI depleted S2 cells and their quantification. DAPI staining is shown in blue, α-tubulin in green. RNAi knockdown experiments were performed by transfecting S2 cells with specific siRNAs against each protein (see “Methods”). Scale bar = 5 μm. Five classes of defects were considered: A multipolar spindle (MS); B chromosome misalignments in metaphase (CM); C chromatin bridges (CB); D long intercellular bridges (LIB); E multinucleated cells (MC); F Quantitative analysis of defects scored in RNAi-treated and control cells (Table 5) is based on the following numbers: at least 100 prometaphases and metaphases for MS, 70 metaphases for CM, 300 telophases for LIB and CB, 5500 interphases for MC. Three independent experiments were performed. G Percent of cells showing a decrease of fluorescence intensity at the midbody in MRG15, Tip60, or YETI depleted cells (black histograms) compared to the mocks (white histograms). The results are based on three independent experiments. H WB showing a decrease of total amount of MRG15, Tip60, and YETI proteins in RNAi-treated S2 cells compared to the mocks; a-tubulin is used as loading control