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

Fig. 1. Nuclear positioning is associated with ER morphology remodeling and requires Climp-63.

Fig. 1

a FIB-SEM representative image of non-LPA stimulated, non-polarized wound-edge U2OS cell. One stack of 998 SEM images from one leading edge cell. b Representative images of perinuclear (yellow insets) and peripheral (green insets) regions. The ER is highlighted in red, light red represents tubules (<1 µm), dark red represents sheets (>1 µm). Measurements represent the depth of the section. c as in a, but an LPA stimulated and polarized cell. One stack of 1798 SEM images from one leading edge cell. d as in b, but an LPA stimulated and polarized cell. e Representative widefield epifluorescence images of wound-edge NIH3T3 fibroblasts. Cells were immunostained for cell-cell contacts (purple; β-catenin), centrosome (white; pericentrin) and DNA (DAPI, blue). f Nuclear positions relative to the cell centroid of cells represented in e. Significance (Two-tailed unpaired t-test) was calculated between experimental condition and the scramble siRNA with LPA. ****p < 0.0001 (Scramble no LPA, Nesprin-2G, Climp-63 #1, Climp-63 #2, Triple KD); *p < 0.05. Box shows first quartile, median and third quartile, and whiskers show 10–90% percentile. Experiments were repeated ≥3, except (a–d). Scale bars: a, b, c, d 1 µm; e 40 µm.