Fig. 2: TRF2 association with Emerin and INM proteins is functional to the establishment of cell polarity. | Cell Death & Disease

Fig. 2: TRF2 association with Emerin and INM proteins is functional to the establishment of cell polarity.

From: TRF2 interaction with nuclear envelope is required for cell polarization and metastasis in triple negative breast cancer

Fig. 2

A Left panel: representative images of nuclear morphology of TRF2-depleted MDA-MB-231 cells visualized by immunofluorescence of Lamin A/C and DAPI (scale bar, 10 μm). Right panel: Box-and-whisker plots (10–90 percentile) of nuclear circularity index (n = 100). B Left panel: representative images of nuclear morphology of MDA-MB-231 cells stable overexpressing TRF2 WT (pTRF2) or TRF2_ΔB (pTRF2_ΔB) deletion mutant and their control (pBabe) visualized by immunofluorescence of Lamin A/C and DAPI (scale bar, 10 μm). Right panel: Box-and-whisker plots (10–90 percentile) of nuclear circularity index (n = 90). C Starved confluent monolayers of stable TRF2 interfered (shTRF2 N1) MDA-MB-436 cells and their respective control (shSCR) were scratch-wounded and stimulated by Lysophosphatidic acid (LPA) (10 μM). Cells fixed at different times upon LPA treatment, were stained with a centrosome marker (pericentrin), Phalloidin, and DAPI. Representative images at 120 min are shown. An example of measurement of pericentrin orientation angle is shown in shTRF2 representative image. For each cell, a first line was drawn from the center of the nucleus to the wound edge and a second line was drawn from the center of the nucleus to the center of the centrosome. The angle formed between the lines was measured. Scale bars, 10 μm. D Quantification of centrosome orientation angle in untreated or LPA treated cells for indicated times and experimental conditions. E Stable TRF2 interfered (shTRF2 N1) MDA-MB-436 cells and their respective control (shSCR) were transfected with indicated siRNA and processed as described in C. Quantification of centrosome orientation angle upon LPA treatment (120 min) in the indicated experimental conditions. F Stable TRF2 overexpressing MDA-MB-436 cells (pTRF2) and respective control cells (pBabe) were processed as described in C. Representative images at 120 min are shown. G Quantification of centrosome orientation angle in untreated or LPA treated cells for indicated times and experimental conditions. H Stable TRF2 overexpressing MDA-MB-436 cells (pTRF2) and respective control cells (pBabe) were transfected with indicated siRNA and processed as described in C. Quantification of centrosome orientation angle upon LPA treatment (120 min) in the indicated experimental conditions. I MDA-MB-436 stable overexpressing TRF2 WT (pTRF2) or TRF2_ΔB (pTRF2_ΔB) deletion mutant and their control (pBabe) were processed as described in C. Representative images at 120 min are shown. H Quantification of centrosome orientation angle in untreated or LPA treated cells for indicated experimental conditions at 120 min. For A, B, Mann Whitney test was used to calculate statistical significance. For (D, E, G, H, J), data are the mean ± SD (n = 3 independent experiments) and two-tailed t student test was used to calculate statistical significance. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

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