Figure 3.
Figure 3. Disruption of actin or LINC complexes decreases chromatin herniation without rescuing lamina disorganization. (A) Representative images of U2OS GFP-NLS shLmnB1 cells arrested with thymidine, treated with indicated drugs, and labeled with DAPI and lamin A antibodies (LmnA). Bot., bottom section of nuclear lamina; Mid., midsection of nucleus rim used for analysis; MIP, maximum intensity projection of 11 and 12 sections with 0.55-µm step size. (B) Quantification of lamina holes in each nucleus rim. n > 160 cells from two experiments. Ns, P > 0.05, Mann–Whitney test. (C) Representative images of U2OS GFP-NLS shLmnB1 cells treated with indicated siRNAs, arrested with thymidine, and labeled with DAPI and LmnA antibodies. MIPs = 13 and 15 sections with 0.55-µm step size. (D) Quantification of lamina holes in each nucleus rim. n > 60 cells from three experiments. Ns, P > 0.05, Mann–Whitney test. (E) MIPs of DAPI-labeled cells arrested with thymidine and treated with indicated drugs (11 and 13 sections with 0.55-µm step size). Arrows indicate chromatin herniations. (F) Quantification of chromatin hernia formed during 24-h time-lapse imaging. Graph represents pooled data (columns) from three experiments (points, n > 120 cells). ***, P < 0.001, Fisher’s exact test. (G) Quantification of maximum area of chromatin herniations present or emerging during 24-h time-lapse imaging. Boxes are quartiles and whiskers are 5th to 95th percentiles. n = 160 DMSO, 22 blebbistatin nuclei from three experiments. *, P < 0.05, Mann–Whitney test. (H) MIPs of DAPI labeled cells arrested with thymidine and treated with indicated drugs (12 and 14 sections with 0.42-µm step size). Arrows indicate chromatin herniations. (I) Quantification of chromatin hernia formed during 24-h time-lapse imaging. Graph represents pooled data (columns) from three experiments (points, n > 80 cells). ***, P < 0.001, Fisher’s exact test. (J) Quantification of maximum area of chromatin herniations present or emerging during 24-h time-lapse imaging. n = 110 control and 33 Sun1 nuclei from three experiments. *, P < 0.05, Mann–Whitney test. Bars, 10 µm.

Disruption of actin or LINC complexes decreases chromatin herniation without rescuing lamina disorganization. (A) Representative images of U2OS GFP-NLS shLmnB1 cells arrested with thymidine, treated with indicated drugs, and labeled with DAPI and lamin A antibodies (LmnA). Bot., bottom section of nuclear lamina; Mid., midsection of nucleus rim used for analysis; MIP, maximum intensity projection of 11 and 12 sections with 0.55-µm step size. (B) Quantification of lamina holes in each nucleus rim. n > 160 cells from two experiments. Ns, P > 0.05, Mann–Whitney test. (C) Representative images of U2OS GFP-NLS shLmnB1 cells treated with indicated siRNAs, arrested with thymidine, and labeled with DAPI and LmnA antibodies. MIPs = 13 and 15 sections with 0.55-µm step size. (D) Quantification of lamina holes in each nucleus rim. n > 60 cells from three experiments. Ns, P > 0.05, Mann–Whitney test. (E) MIPs of DAPI-labeled cells arrested with thymidine and treated with indicated drugs (11 and 13 sections with 0.55-µm step size). Arrows indicate chromatin herniations. (F) Quantification of chromatin hernia formed during 24-h time-lapse imaging. Graph represents pooled data (columns) from three experiments (points, n > 120 cells). ***, P < 0.001, Fisher’s exact test. (G) Quantification of maximum area of chromatin herniations present or emerging during 24-h time-lapse imaging. Boxes are quartiles and whiskers are 5th to 95th percentiles. n = 160 DMSO, 22 blebbistatin nuclei from three experiments. *, P < 0.05, Mann–Whitney test. (H) MIPs of DAPI labeled cells arrested with thymidine and treated with indicated drugs (12 and 14 sections with 0.42-µm step size). Arrows indicate chromatin herniations. (I) Quantification of chromatin hernia formed during 24-h time-lapse imaging. Graph represents pooled data (columns) from three experiments (points, n > 80 cells). ***, P < 0.001, Fisher’s exact test. (J) Quantification of maximum area of chromatin herniations present or emerging during 24-h time-lapse imaging. n = 110 control and 33 Sun1 nuclei from three experiments. *, P < 0.05, Mann–Whitney test. Bars, 10 µm.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal