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. 2022 May 19;13:2763. doi: 10.1038/s41467-022-30388-3

Fig. 2. ER accumulates at the perinuclear region during nuclear positioning and requires Climp-63.

Fig. 2

a Representative point-scanning confocal Airyscan images of wound-edge GFP-KDEL-expressing fibroblasts treated with scramble or Climp-63 siRNA, stimulated and non-stimulated with LPA. Perinuclear region of interest (ROI) are highlighted (yellow box) and represented as grey scale (top) and as heatmap (bottom). b Quantification of perinuclear ER accumulation of cells represented in a. treated with scramble (grey) or Climp-63 (green) siRNA. Significance (Two-tailed unpaired t-test) was calculated between experimental conditions and scramble with LPA (second condition). c Fluorescence spinning-disk confocal kymograph of nuclear movement and perinuclear ER accumulation in wound-edge GFP-KDEL-expressing cells treated with scramble (top) or Climp-63 (bottom) siRNA upon LPA stimulation. Images correspond to the maximal intensity projection of z-stacks from ventral to dorsal side of the cell. The signal intensity is represented as a thermal heatmap. d Quantification of perinuclear ER accumulation in LPA stimulated cells treated with scramble (grey, n = 22 cells of 4 independent experiments) or Climp-63 (green, n = 21 cells of 4 independent experiments) siRNA, over time, represented in c. Significance (Two-tailed unpaired t-test) was calculated between scramble and Climp-63 siRNAs for each time point. e Quantification of perinuclear ER accumulation (grey line) and the nuclear movement over time (blue line), in LPA stimulated cells in scramble (left, n = 61 cells of 4 independent experiments) or Climp-63 (right, n = 37 cells of 4 independent experiments) siRNA. Perinuclear ER accumulation is the same data as in d. Scale bars: a, c 10 µm. Error bars, SEM. **p < 0.01, *p < 0.05.