Fig. 3. Perinuclear ER accumulation depends on Climp-63 luminal domain but it is independent of microtubules.

a Schematic representation of Climp-63 synthetized plasmids. Climp-63 is the full-length protein. The cytoplasmic (Cyto) mutant lacks the luminal domain. The luminal (Lum) mutant lacks the cytoplasmic domain. The (−) MT mutant does not bind to microtubules. The (+) MT mutant is constitutively bound to microtubules. The asterisks represent the siRNA resistant mutations. b Average nuclear positions relative to the cell centroid of cells treated with scramble or Climp-63 siRNA and microinjected with the indicated plasmids. Significance (Two-tailed unpaired t-test) was calculated between the experimental conditions and Climp-63 siRNA cells (second condition). ****p < 0.0001 (Scramble alone, Climp-63); Box show first quartile, median and third quartile, and whiskers show 10–90% percentile. c Quantification of the percentage of cells with perinuclear ER accumulation. Significance (Two-tailed unpaired t-test) was calculated between experimental conditions and Climp-63 expressing cells (third condition). Error bars, SEM. d Representative point-scanning confocal images of wound-edge cells treated with Climp-63 siRNA and microinjected with the plasmids shown in a. Perinuclear ROI are highlighted (yellow box) and represented as grey scale (top) and as heatmap (bottom). Experiments were repeated ≥3. Scale bars: d 10 µm. **p < 0.01, *p < 0.05.