Files in this Data Supplement:
Fig. S1. LUMA is highly conserved. ClustalW alignment of a representative selection of orthologous LUMA sequences. Identical residues are displayed on a red background, similar amino acids are shown as red letters in boxes. The predicted TMD sequences for the human orthologue are indicated as TM 1 to TM 4.
Fig. S2. ClustalW alignment of selected orthologous LUMA sequences with murine LUMA. Phaeodactylum tricornutum (algae), Arabidopsis thaliana, Caenorhabditis elegans, Solanum tuberosum (potato), Desulfitobacterium hafniense (bacterium), and Silicibacter sp. Tm1040 (bacterium). Identical residues are displayed on a black background, similar amino acids are shown as bold letters in boxes.
Fig. S3. Secondary structure prediction for LUMA. Individual predictions and consensus of the secondary structure prediction for LUMA obtained by GOR1, NNPredict, HNN, SOPM, and SOPMA. Grey: α-helix, black: β-sheet. The predicted arrangement of membrane-spanning sequences is indicated between the consensus and the individual predictions. ‘Consensus’ represents an average prediction
Fig. S4. Overexpression of full-length LUMA 1-400 and of LUMA mutants dominantly negative affects the NE structure. LUMA variants were overexpressed in HeLa cells and visualized by immunofluorescence with antibodies against C-terminal tags (LUMA 1-400: anti-V5 antibody; LUMA mutants: anti-Myc antibody). Nuclear envelope was stained with an anti-emerin antibody, chromatin with DAPI. ‘Overview’, ‘LUMA x Emerin’: image overlay of LUMA (green) and emerin (red), Scale bars, 5 m.
Fig. S5. Characterization of the anti-LUMA antibody. (A) Murine neuroblastoma N2a cell homogenate was separated by SDS-PAGE, blotted onto nitrocellulose and probed for LUMA with affinity-purified anti-LUMA antibody. 1: control; 2: serum before affinity purification; 3: affinity-purified anti-LUMA antibody, diluted 1:1000; 4: diluted 1:2000; diluted 1:10,000: (B) Immunofluorescence staining of murine NIH-3T3 cells with the affinity-purified anti-LUMA antibody (LUMA). Control: secondary antibody only; 1:1000, 1:500, 1:100: dilutions of the affinity-purified anti-LUMA antibody. DAPI: chromatin.
Adobe PDF