Picture from Asencio C et al. (2012) Cell "Coordination of kinase and phosphatase activities by Lem4 enables nuclear ...."

(C) control(RNAi), lem-4L(RNAi), and lem-4L(ax475) embryos grown at 16 or 25C were stained with anti-LEM-4L (green) and mAb414 (red). Scale bar, 10 um.

Picture from Dittrich CM et al. (2012) PLoS One "LEM-3 - A LEM domain containing nuclease involved in the DNA damage response in C. ...."

Figure S2 YFP::LEM-3 expression pattern. (A) The transgene opIs383 [Pnpp-1::YFP::lem-3::39UTRlem-3] rescues the hypersensitivity of lem-3(op444) mutants. Data represent the average of three experiments 6 S.D. The progeny of 10 worms were analysed for each experiment. (B) Representative DIC and fluorescence images of embryos expressing YFP::LEM-3; in green: YFP::LEM-3, in red: membrane staining with the dye FM4-64.

Picture from Gonzalez-Aguilera C et al. (2014) Genome Biol "Genome-wide analysis links emerin to neuromuscular junction activity in ...."

Figure S5. Differential expression of EMR-1 and LEM-2. Live imaging of animals expressing EMR-1::GFP and LEM-2::mCherry from single-copy transgenes and under control of the emr-1 and lem-2 promoter, respectively. Both genes are expressed in the NE in most if not all cells, but with considerable variation between tissues as reflected by different intensities in the two upper panels. In the merged image green and magenta represent high and low EMR-1::GFP to LEM- 2::mCherry signal ratio, respectively. Note that these ratios reflect arbitrary fluorescence values and do not necessarily correspond to EMR-1 being more or less abundant than LEM-2.

Picture from Asencio C et al. (2012) Cell "Coordination of kinase and phosphatase activities by Lem4 enables nuclear ...."

Figure 1. Depletion of Worm LEM-4L or Human Lem4 Results in Abnormal Nuclear Morphology (A) Wild-type, lem-4L(ax475), and rescued lem-4L(ax475) embryos were grown at 16 or 25C and stained with anti-LMN-1 (green), mAb414 (red), or Hoechst DNA stain (blue). Scale bar, 10 um. (B) control(RNAi), lem-4L(RNAi), vrk-1(RNAi), and lem-4L(ax475) embryos were grown at 16 or 25C and processed for western blotting with anti-LEM-4L or anti-a-tubulin. Asterisk, cross-reacting band. (C) control(RNAi), lem-4L(RNAi), and lem-4L(ax475) embryos grown at 16 or 25C were stained with anti-LEM-4L (green) and mAb414 (red). Scale bar, 10 mm. (D) HeLa cells were transfected with control siRNA or with either Lem4 oligo 1 or Lem4 oligo 2 siRNA. After 48 hr, cells were costained (Lem4, mAb414) or stained with the indicated antibodies. (E) HeLa cells stably expressing H2B-mCherry (top row) or H2B-mCherry and Lem4-myc (bottom row) were transfected with control, Lem4 oligo 1, or 2 siRNAs. After 48 hr, both cell lines were stained with anti-myc (green). H2B-mCherry was used to visualize DNA (red). (F) The percentage of cells (average of four independent experiments, nR100 cells per experiment) displaying abnormal nuclear shape after RNAi treatment was determined visually. Error bars represent SD. (G) In parallel, cells were resuspended in SDS-PAGE sample buffer and western blotted by using affinity-purified Lem4 antibody. Lower band, truncated Lem4-myc. GAPDH, loading control.

Picture from Asencio C et al. (2012) Cell "Coordination of kinase and phosphatase activities by Lem4 enables nuclear ...."

(C) Embryo expressing LEM-4L-GFP. Upper panel: GFP. Lower panel: DIC. Arrows point to periphery of daughter nuclei after the first embryonic division. Scale bar, 10 mm. (

Picture from Asencio C et al. (2012) Cell "Coordination of kinase and phosphatase activities by Lem4 enables nuclear ...."

Figure S2. Characterization of C. elegans LEM-4L and H. sapiens Lem4, Related to Figure 1 (A and B) Wild-type and lem-4-4L(ax475) embryos grown at 16 or 25C were stained with anti-NUP96 (A) or anti-NUP107 (B), mAb414 and Hoechst. Scale bar, 10 mm. (C) Embryo expressing LEM-4L-GFP. Upper panel: GFP. Lower panel: DIC. Arrows point to periphery of daughter nuclei after the first embryonic division. Scale bar, 10 mm. (D) Western blot using affinity purified Lem4 antibodies #1 and #2. Lem4 antibody #1 was used for all experiments. (E) Digitonin / Triton X-100 permeabilization. HeLa Kyoto cells were fixed and permeabilized with digitonin to permeabilize the plasma membrane (top row) or digitonin + Triton X-100 to permeabilize both the plasma membrane and the NE (bottom row). Cells were stained with antibodies raised against Lem4, ERGIC 53 (lumenal epitope), lamin A (nuclear epitope) and Sec31A (cytosolic epitope). (F) HeLa Kyoto cell transiently transfected with Lem4 1-938::pEGFP-N3, fixed and stained with mAb414 antibody to visualize NPCs and DAPI to visualize DNA. Scale bar: 10 mm. Transiently transfected Lem4 1-938-GFP appears to be full length by western blotting (data not shown). (G) Western blot of cells from Figure 1D. Affinity-purified Lem4 antibody was used to visualize Lem4 depletion efficiency. GAPDH was a loading control. (H) Diagram of truncated Lem4-myc (Figures 1E and 1F). Domains were predicted by SMART. Numbers denote amino acid positions.