Fig. 3

L-DL is an essential structural component of nuclear speckle. a Control GST (CTRL-GST) and GST tagged L-DL (GST-L-DL) were introduced into 293T cells, followed by pull-down with GST agarose beads. L-DL associated proteins were subjected to silver staining, black square denoted bands were subjected to mass spectrometry analysis. b Cellular component terms in the gene ontology (GO) analysis for L-DL associated proteins identified in mass spectrometry. c Representative immunofluorescence images of L-DL (red) and SC35 (green), L-DL (red) and SON (green), L-DL (red) and RBM25 (green), and DAPI (blue) in 293T cells. Scale bars: 10 μm. d Representative immunofluorescence images of L-DL (red) and NPM1 (green), L-DL (red) and PML (green), L-DL (red) and SMN (green), and DAPI (blue) in 293T cells. Scale bars: 10 μm. e CTRL-GST and GST-L-DL were introduced into 293T cells, followed by pull-down with GST agarose beads. L-DL associated proteins were subjected to immunoblotting with anti-SC35, anti-RBM25, anti-NPM1 and anti-PML antibodies. f CTRL-GST and GST-SC35 were introduced into 293T cells, followed by pull-down with GST agarose beads. SC35 associated proteins were subjected to immunoblotting with anti-L-DL antibody. g Left: representative immunofluorescence images of L-DL (red), SC35 (green) and DAPI (blue) in control and L-DL knockout (KO) 293T cells. Scale bars: 10 μm. Right: Densitometric analyses of SC35 fluorescence density. h Left: representative immunofluorescence images of L-DL (red), SON (green) and DAPI (blue) in control and L-DL KO 293T cells. Scale bars: 10 μm. Right: densitometric analyses of SON fluorescence density. Statistical analysis was performed using two-tailed Student’s t-test; *** P < 0.001; error bars denote SEM