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. 2010 Dec 20;108(1):131–136. doi: 10.1073/pnas.1000824108

Fig. 4.

Fig. 4.

Effect of Lamin A/C on actin cables and TAN lines (A) Panels from movies of Lmna+/+ (Upper) and Lmna−/− (Lower) MEFs expressing Lifeact-mCherry showing dorsal actin cables near the nucleus. The wound edge is located at the top, yellow circles denote nuclei, and arrows denote actin cables that move either with (Upper) or over (Lower) the nucleus. Time is in minutes after LPA stimulation. (Scale bar, 5 μm.) (B) Velocities of nuclei and actin cables from the cells described in A. Error bars are SEM from three experiments and 45 total cells. Nuclear velocity in LMNA−/− MEFs was significantly different from that in Lmna+/+ MEFs (P < 0.01; there was no significant difference in the velocity of actin cables). (C) Relative intensity of nesprin-2G immunofluorescence compared with nonexpressing neighboring cells. Values are the percentage difference from nonexpressing neighboring cells. Error is SEM for >30 cells from three separate experiments. Nesprin-2G intensity was significantly different (P < 0.05) for all variants that cause EDMD or DCM, except T528K, compared with WT- or FPLD-causing variants. (D) Nucleus (red) and centrosome (blue) positions in nesprin 2G-depleted NIH 3T3 fibroblasts and Lmna−/− MEFs expressing GFP-mininesprin-2G. Centrosome orientation (percentage of cells) is located below the histogram. Error bars are SEM for >45 cells from at least three independent experiments. Centrosome orientation and nuclear position in nesprin 2G-depleted cells expressing GFP-mininesprin-2G were significantly different from those in nonexpressing cells (P < 0.01). (E) Immunofluorescence images of nuclei from a Lmna+/+ MEF (Upper) and Lmna−/− MEF (Lower) expressing GFP-mininesprin-2G (green in merge) and stained for endogenous SUN2 (red in merge). Arrows indicate colocalized nesprin-2G and SUN2. (Scale bar, 5 μm.). (F) Percentage of GFP-mininesprin-2G TAN lines that persist >20 min. Error bars are SEM from >40 cells and three independent experiments. The differences between Lmna+/+ and Lmna−/− MEFs and between GAPDH-depleted and SUN2-depleted NIH 3T3 fibroblasts were significant (P < 0.05).