Fig. 1: Elys-KD in S2 cells does not lead to a notable loss of NPCs at the NE.

a, b, d Immunostaining of control, Lam-KD, Elys-KD, or (Lam-KD plus Elys-KD) S2 cells with anti-Elys, anti-Lamin and Mab414 antibodies (a), with anti-Elys and Mab414 antibodies (b), or with anti-LBR, anti-Elys and anti-Lamin antibodies (d). Scale bars 1 µm (a, d), 10 µm (b). c ImageJ quantification of Mab414 average fluorescence intensity (normalized on average Dapi fluorescence) across the NE in Elys-KD (two biological replicates, n = 75) and control (two biological replicates, n = 75) S2 cells. The ratio of median values is indicated. P-value was estimated in an M-W U-test. e Western-blot analysis of proteins, co-immunoprecipitated with anti-Elys or anti-Nup107 antibodies from S2 extracts, probed by anti-Elys, or anti-Nup107 antibodies (n.i. – non-immune serum, IP/input ratio 1:4.5). Note that the spliced bands ran in parallel on the same gel. The sizes of molecular weight markers are indicated to the right. f, g Immunostaining of S2 cells with anti-CenpA (kinetochores, violet), anti-Elys (red), anti-α-Tubulin (green) in metaphase (f), with anti-Elys (red), anti-Mab414 (green), anti-LBR (violet), Dapi (blue) in anaphase (g). Scale bars 1 µm (f, g). Arrows point to the Elys concentrated around decondensing chromatin (g). h Immunostaining of control and Elys-KD S2 cells with Mab414 antibodies (green) counterstained with Dapi (blue) in telophase. Scale bar 1 µm.