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. 2012 Jul 23;7(7):e41537. doi: 10.1371/journal.pone.0041537

Figure 9. Negr1-I87N is a loss-of-function mutation.

Figure 9

A–C. NSC-34 cells expressing EGFP alone (control) or together with Negr1-WT or Negr1-I87N mutants at 0 min (t1) and following 60 min (t2) of cell aggregation. D. Histogram showing cell aggregation expressed as the ratio of 0 min (t1) and 60 min (t2) time points. EGFP+empty vector (control): 1.06±0.05; NEGR1-WT+EGFP: 2.11±0.2; EGFP+NEGR1-I87N: 0.93±0.2. Data represent means ± sd calculated from three independent experiments. E–G. Confocal images showing hypothalamic neurons immunostained for the neuronal marker βIII-tubulin (red) cultured together with transfected NSC-34 cells (green). NSC34 cells were transfected with (E) pEGFP together with an empty pcDNA-vector (control), (F) pEGFP+pcDNA3-NEGR1-WT and (G) EGFP+NEGR1-I87N. H. Mean neurite lengths of hypothalamic neurons relative to control (set to 100%). Error bars represent SEM from three independent experiments (∼400 neurites per condition). Two-tailed Student's test; *P<0.05; **P<0.01; *** P<0.001. Scale: 100 µm (A), 20 µm (E).