Fig 1: SUN1 and SUN2 influence isolated VSMCs spreading. (A) Representative confocal immunofluorescence microscopy images of rhodamine phalloidin (red), SUN1 or SUN2 (green), and DAPI (blue) stained VSMCs grown on 12 kPa hydrogels. Graphs show IF analysis of (B) cell area, (C) nuclear area, and (D) nuclear area:cytoplasmic area ratio of control, SUN1- and SUN2-depleted VSMCs. Graphs represent combined data of three independent experiments analysing >300 cells per group (* p < 0.05 and ** p < 0.01).
Fig 2: Telomere clustering occurs at the mid-late pachytene stage in Ythdc2iKO spermatocytes.(A) Immunofluorescence of CREST and SUN1 in surface nuclear spreads of pachytene cells.(B) Immunofluorescence of SUN1 in pachytene cells prepared from testicular single cell suspension. See also Videos S1 and S2.(C–D) Substaging of pachytene cells by the H1t expression. H1t is absent in early pachytene cells, appears in mid-pachytene cells, and is abundant in late pachytene cells (C). Inset (C) shows the H1t (black and white) signal in the entire nucleus. The plot (D) shows the percentage (mean ± s.d.) of each type of pachytene spermatocytes. Scale bars, 10 μm.
Fig 3: Nuclear shape, internuclear distances, and LINC complex proteins were unaffected in TA muscles from nesprin-1ΔCH mice. (A) TA skeletal muscle fibers isolated from either WT or nesprin 1ΔCH (N1ΔCH) mice were fixed and stained using Lamin A/C (A/C), Lamin B1 (B1), Sun1, Sun2 (green), and α-actinin (red) antibodies. Arrowheads denote the specific localization to the NE. (B) Western blots of the LINC complex proteins Sun1 and Sun2, the LINC-associated protein emerin, and the nuclear lamins A/C, B1, and B2 from WT and nesprin 1ΔCH (KO) TA muscle lysates. Note that the localization and levels were unaffected in nesprin 1ΔCH compared with WT controls. GAPDH served as a loading control. (C) Internuclear distances were quantified in TA muscle fibers isolated from WT (blue bars) and nesprin 1ΔCH (green bars) mice shown in A. Note that the distances between nuclei were unaffected. n = 176–205 and n = 140–230 internuclear distances were counted for nesprin 1ΔCH and WT, respectively. (D) Low-magnification representative images of those shown in A. n = at least three to four pups per genotype for each panel. Bars: (A) 5 µm; (D) 10 µm. DIC, differential interference contrast.
Fig 4: SUN2 KO VSMCs display reduced spreading. (A) WB of wild-type (WT) and SUN2 KO aortic samples. Each lane corresponds to an independent aortic sample isolated from different WT and SUN2 KO mice. (B) qPCR analysis of SUN1 and SUN2 mRNA expression in WT aortae. (C) Representative images of isolated WT and SUN2 KO mouse aortic VSMCs stained for F-actin (red), SUN2 (green), and DAPI (blue). Graphs show (D) cell area, (E) nuclear area, and (F) nuclear area:cell area ratio of WT and SUN2 KO VSMCs. Graphs represent the combined data of three independent experiments analysing >200 cells per group (** p < 0.01 and *** p < 0.0001).
Fig 5: The LINC complex in mRNA export. (A) Efficiency of Nesprin-2 knockdown (KD) by shRNA treatment in HeLa cells is shown by immunofluorescence with Nesprin-2 specific antibodies pAbK1 and by western blot analysis. The quantitative determination of the knockdown is depicted in a bar graph (left). Scale bar, 5 µm. (B) RNA FISH analysis of Nesprin-2 depleted cells as shown by immunofluorescence analysis. Left side, distribution of the N/C intensity range. (C) The mean nuclear/cytoplasmic (N/C) intensity ratio of control and Nesprin-2 KD cells. The values represent the mean ± SD of more than 100 cells analyzed. (D) Immunofluorescence analysis of GFP-SUN1-?SUN expressing cells. Nesprin-2 was detected by pAbK1. Scale bar, 10 µm.
Supplier Page from Abcam for Anti-SUN1 antibody