![Figure 4. LINC complexes contribute to the formation of PNEIs. (A) Representative nuclei showing varying levels of PNEI observed upon the indicated treatments. (B–E) Nuclei of early prophase cells monitored by live-cell imaging were scored for PNEI. Nuclei were counted as PNEI positive if deep nuclear invaginations appeared before NEBD. Bar plots represent the percentage of nuclei with PNEI upon RNAi or the addition of cytoskeletal inhibitors (five to six experiments; means ± SEM; n ≥ 14 per condition). (F) PNEI analysis derived from videos of cells expressing CFP, CFP-GCN4-SUN1 (DN-SUN1), CFP-GCN4-SUN2 (DN-SUN2), or CFP-GCN4-SUN2(H628A) in the lumen of the ER 48 h after transfection (five experiments; means ± SEM; n ≥ 12 per condition). (G) PNEI analysis derived from videos of reporter cells expressing cytoplasmic (cyt) CFP, CFP-NES2c (wild type [wt]; DN-KASH2), or C-terminally extended (ext) CFP-NES2c 48 h after transfection (seven experiments; means ± SEM; n ≥ 27 per condition). (bottom) CFP fluorescence of the respective proteins is shown. Bars, 5 µm.](https://cdn.rupress.org/rup/content_public/journal/jcb/204/7/10.1083_jcb.201310116/5/jcb_201310116_fig4.jpeg?Expires=1771608228&Signature=ApyhIEq5rFuOZwcvSaMqHBd29dVP5pd0KgdtyCEosyZlWNToBjEMrr8TgVS4mNWlCO5gvYVKyqbFmrXiFq7-BeY33xIWdhkRFT~rdOOoYDupdbUeU2b3FT6g2ojVXfGaED6TnLZ9JztQ~gkVlU~bxwno80laE22TV9rc12wQVICJfgVjs6Txv56GpTHdSI81dFS3elzcsQ0ZwVoBSU-KSZhtRFZdpl8X01mbQfXhffhYCxe0YzRioF~39pDhAyWsJHGDDEpZiSls~gebRQVNmj2mCTHUNlhZ0LkdT5R0q03sv5PO8T19gLwaz7XlIPFN1DkP708fRv2E4TwtJ~n6qA__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
LINC complexes contribute to the formation of PNEIs. (A) Representative nuclei showing varying levels of PNEI observed upon the indicated treatments. (B–E) Nuclei of early prophase cells monitored by live-cell imaging were scored for PNEI. Nuclei were counted as PNEI positive if deep nuclear invaginations appeared before NEBD. Bar plots represent the percentage of nuclei with PNEI upon RNAi or the addition of cytoskeletal inhibitors (five to six experiments; means ± SEM; n ≥ 14 per condition). (F) PNEI analysis derived from videos of cells expressing CFP, CFP-GCN4-SUN1 (DN-SUN1), CFP-GCN4-SUN2 (DN-SUN2), or CFP-GCN4-SUN2(H628A) in the lumen of the ER 48 h after transfection (five experiments; means ± SEM; n ≥ 12 per condition). (G) PNEI analysis derived from videos of reporter cells expressing cytoplasmic (cyt) CFP, CFP-NES2c (wild type [wt]; DN-KASH2), or C-terminally extended (ext) CFP-NES2c 48 h after transfection (seven experiments; means ± SEM; n ≥ 27 per condition). (bottom) CFP fluorescence of the respective proteins is shown. Bars, 5 µm.
