Fig 1: The density of LINC complexes is enriched at the sites of nuclear lamina indentations.(A) SIM images of LINC complexes labeled with Syne-2 antibody (red) in a deformed nucleus. A focal plane in SIM images located in the central zone was selected to show (B) LINC complexes fluorescence signals at the bottom of (C) lamin indentations sites. (D) The merged image of lamin and LINC complexes from the same focal plane showed that both signals spatially co-localized. (E) Apical actin filaments and (F) LINC complexes observed in SIM showed (G) a strong co-localization on the merged image. (H) Schematic representation of the spatial organization of actin microfilaments (green), nuclear lamina (yellow) and LINC complexes (red) in a rectangular-shaped endothelial cell. (I) Evolution of the normalized density of LINC complexes for rounded and non-indented nuclei (white bar, n = 12), for non-indented elongated nuclei (dashed bar, n = 11) and for elongated nuclei with deep indentations (black bar, n = 14). Bars are mean ± SD. *p < 0.05, unpaired Student's test. Scale bars correspond to 1 µm.
Fig 2: Immunohistochemical staining for SUN1, SUN2, and nesprin-2. Specimens of breast tumor were stained using pAbs against SUN1 (A), SUN2 (B), and nesprin-2 (C). Representative cases including noncancerous (left) and cancerous regions (middle and right) are shown. The upper panels were obtained at lower magnification than the lower panels. Bar, 50 μm.
Fig 3: mRNA expression of linker of nucleoskeleton and cytoskeleton (LINC) complex and nuclear lamina components was also reduced in cancer. (A) SUN1, SUN2, nesprin-2, and lamin A/C mRNA expression in human mammary epithelial and breast tumor tissues was measured by real-time PCR. Results are presented as means ± SD. (B) SUN1, SUN2, nesprin-2, and lamin A/CmRNA levels in normal mammary epithelial cells (MCF10A), human breast tumor cells (MDA-MB-231 [described as MM231], MCF7; and ZR75-30), and human cervical cancer cells (HeLa) were analyzed by real-time PCR. The values were normalized against GAPDH mRNA. Results are presented as means ± SD.
Fig 4: Loss of lamin A/C, SUN1, SUN2, and nesprin-2 staining in the cancerous regions of breast cancer tissue. (A–D) Noncancerous and cancerous regions of breast cancer tissue were stained with anti-lamin A/C (A), anti-SUN1 (B), anti-SUN2 (C), and anti-nesprin-2 (D) pAbs; representative staining patterns are shown. The upper panels were obtained at lower magnification than the lower panels. Most myoepithelial cells stained more strongly for anti-SUN1 pAbs in cancer-associated noncancerous regions than in most mammary epithelial cells, but not in all cases (see Fig.2).
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