Journal: bioRxiv

Article Title: Nucleoporin Elys attaches peripheral chromatin to the nuclear pores in interphase nuclei

doi: 10.1101/2023.08.16.553518

Figure Lengend Snippet: ( A , B , D ) Immunostaining of control, Lam-KD, Elys-KD, or (Lam-KD plus Elys-KD) S2 cells with anti-Elys, anti-Lam and Mab414 antibodies ( A ), with anti-Elys and Mab414 antibodies ( B ), or with anti-LBR, anti-Elys and anti-Lam 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 replicates, n = 75) and control (two replicates, n = 75) S2 cells. P value was estimated in a 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). ( 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 Elys-KD S2 cells with Mab414 antibodies (green) counterstained with Dapi (blue) in telophase. Scale bar 1 µm.

Article Snippet: As the primary, mouse monoclonal anti-Lam [ ] (ADL84, 1:500), guinea pig polyclonal anti-LBR [ ] (1:1000), mouse monoclonal Mab414 (1:300; Abcam ab24609), chicken polyclonal anti-CenpA [ ] (1:600), rabbit polyclonal α-Tubulin (1:2000; Abcam ab18251) and rabbit polyclonal anti-Elys [ ] (1:1000) antibodies were used.

Techniques: Immunostaining, Control, Fluorescence, Western Blot, Immunoprecipitation